Cosmetic formulation comprising 3-isothiazolones

ABSTRACT

Disclosed herein are certain novel compounds, which are most properly designated as 3-isothiazolones. These compounds and compositions containing them exhibit a broad spectrum of biocidal properties and are particularly effective for the control of microorganisms.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a division of application Ser. No. 910,730, filedMay 30, 1978, which was a division of Ser. No. 368,168, filed June 8,1973, now U.S. Pat. No. 4,105,431, which is a continuation-in-part ofSer. No. 836,660, filed June 25, 1969, now U.S. Pat. No. 3,761,488,which is a continuation-in-part of Ser. No. 672,437, filed Oct. 3, 1967,now U.S. Pat. No. 3,523,121, which is a continuation-in-part of Ser. No.621,780, filed Mar. 9, 1967, now abandoned.

This invention relates to novel substituted 3-isothiazolones, saltsthereof, their preparation, agricultural compositions containing then,and their utilization in the control of living organisms.

These novel 3-isothiazolones (hereinafter referred to at times as"isothiazolones") are represented by the formula ##STR1## wherein Y isan unsubstituted or substituted alkyl, alkenyl, or alkynyl group of 1 to18 carbon atoms, preferably 4 to 18 carbon atoms, an unsubstituted orsubstituted cycloalkyl group having a 3 to 12 carbon atom ring, anunsubstituted or substituted aralkyl group of up to 10 carbon atoms, oran unsubstituted or substituted aryl group of up to 10 carbon atoms,

R is hydrogen, halogen, or a (C₁ -C₄)alkyl group, and

R' is hydrogen, halogen, or a (C₁ -C₄)alkyl group, provided that when Yis methyl or ethyl then both R and R' may not be hydrogen.

By a substituted alkyl group is meant an alkyl group having one or moreof its hydrogen atoms replaced by another substituent group. Examples ofthe substituted alkyl groups which characterize 3-isothiazolones of thisinvention include hydroxyalkyl, haloalkyl, cyanoalkyl, alkylaminoalkyl,dialkylaminoalkyl, arylaminoalkyl, carboxyalkyl, carbalkoxyalkyl,alkoxyalkyl, aryloxyalkyl, alkylthioalkyl, arylthioalkyl,haloalkoxyalkyl, cyclo laminoalkyl, such as morpholinoalkyl,piperidinoalkyl, pyrrolidonylalkyl, and the like, carbamoxyalkyl,alkenyl, haloalkenyl, alkynyl, haloalkynyl, isothiazolonylalkyl, and thelike.

By a substituted aralkyl group is meant an aralkyl group having one ormore of the hydrogen atoms on either the aryl ring or the alkyl chainreplaced by another substituent group. Examples of the substitutedaralkyl groups which characterize 3-isothiazoles of this inventioninclude halogen, lower alkyl-, or lower alkoxy-substituted aralkylgroups, and the like.

By a substituted aryl group is meant an aryl group, such as benzene,naphthalene, or pyridine, having one or more of the hydrogen atoms onthe aryl ring replaced by another substituent group. Examples of suchsubstituent groups include halogen, nitro, lower alkyl, loweralkylacylamino, lower carbalkoxy, sulfamyl, and the like.

The isothiazolones described above can form novel acid salts which alsoexhibit biocidal activity. Preparation of these biocidally active saltsis readily achieved by reacting the above designated 3-isothiazoloneswith a strong inorganic or organic acid. Typical strong acids includehydrochloric, nitric, sulfuric, hydrobromic, chlorosulfuric,chloroacetic, oxalic, maloic, succinic, p-toluenesulfonic, and the like.Separation of the acid salts from the reaction medium is accomplished byany convenient means known to one skilled in the art.

Representative Y substituents include methyl, ethyl, propyl, isopropyl,butyl, hexyl, octyl, decyl, pentadecyl, octadecyl, cyclopropyl,cyclohexyl, benzyl, 3,4-dichlorobenzyl, 4-methoxybenzyl, 4-chlorobenzyl,3,4-dichlorophenyl, hydroxymethyl, chloromethyl, chloropropyl,diethylaminoethyl, cyanoethyl, carbomethoxyethyl, ethoxyethyl,2-methoxy-1-bromoethyl, 3,3,5-trimethylcyclohexyl, phenoxyethyl,p-chloroanilinomethyl, phenylcarbamoxymethyl, allyl, propynyl, vinyl,carboxyethyl, 1-isothiazolonylethyl, and 1,2,2,-trichlorovinyl.

Representative R substituents include hydrogen, bromo, chloro, iodo,methyl, ethyl, propyl, isopropyl, butyl and t-butyl.

Representative R' substituents are hydrogen, chloro, bromo, iodo,methyl, ethyl, propyl, isopropyl, butyl, t-butyl, chloromethyl,chloropropyl, bromomethyl, bromoethyl and bromopropyl.

Typical compounds within the scope of Formula I include the following:

2-propyl-3-isothiazolone,

2-butyl-3-isothiazolone,

2-t-octyl-3-isothiazolone,

2-decyl-3-isothiazolone,

2-octyldecyl-3-isothiazolone,

2-cyclohexyl-3-isothiazolone,

4-chloro-2-methyl-3-isothiazolone,

4-bromo-2-methyl-3-isothiazolone,

5-chloro-2-methyl-3-isothiazolone,

5-chloro-2,4-dimethyl-3-isothiazolone,

4-bromo-5-chloro-2-methyl-3-isothiazolone,

4-bromo-2-cyclohexyl-3-isothiazolone,

4,5-dichloro-2-ethyl-3-isothiazolone,

4-methyl-2-octyl-3-isothiazolone,

4,5-dimethyl-2-octyl-3-isothiazolone,

2-benzyl-3-isothiazolone,

2-benzyl-4,5-dichloro-3-isothiazolone,

2-benzyl-5-chloro-3-isothiazolone,

2-(2',4'-dichlorobenzyl)-3-isothiazolone

2-(4'-methoxybenzyl)-3-isothiazolone

2-(4'-ethylbenzyl)-3-isothiazolone

2-(3',4'-dichlorophenyl)-3-isothiazolone,

2-(3',4'-dichlorophenyl)-4-methyl-3-isothiazolone,

2-(2-cyanoethyl)-3-isothiazolone,

2-(2-carbomethoxyethyl)-3-isothiazolone,

2-carbomethoxymethyl-3-isothiazolone,

2-(2-ethoxyethyl)-3-isothiazolone,

2-(3',3',5'-trimethylcyclohexyl)-3-isothiazolone,

2-(2-phenoxyethyl)-3-isothiazolone,

2-(2-methoxyethyl)-3-isothiazolone,

2-(3',4'-dichloroanilinomethyl)-3-isothiazolone,

2-(4'-chloroanilinomethyl)-3-isothiazolone,

2-(4'-nitroanilinomethyl)-3-isothiazolone,

2-morpholinomethyl-3-isothiazolone,

2-piperidinomethyl-3-isothiazolone,

2-phenylcarbamoxymethyl-3-isothiazolone,

2-(3'-chlorophenylcarbamoxymethyl)-3-isothiazolone,

2-(3',4'-dichlorophenylcarbamoxymethyl)-3-isothiazolone,

2-allyl-3-isothiazolone,

2-propynyl-3-isothiazolone,

2-vinyl-3-isothiazolone,

5-chloro-2-vinyl-3-isothiazolone,

2-methoxymethyl-3-isothiazolone,

2-(2-carboxyethyl)-3-isothiazolone,

2-(2-carb-n-butoxyethyl)-3-isothiazolone,

2-[1-(N-pyrrolidonyl)ethyl]-3-isothiazolone,

2-[1-(N-isothiazolonyl)ethyl]-3-isothiazolone,

2-(1,2,2-trichlorovinyl)-3-isothiazolone,

2-(1-bromo-2-methoxyethyl)-3-isothiazolone,

2-(2-chloroethyl)-3-isothiazolone,

2-(3-chloropropyl)-3-isothiazolone,

2-cyclopropyl-3-isothiazolone,

2-[2-(4'-chlorophenyl)ethyl]-3-isothiazolone,

2-n-hexyl-3-isothiazolone,

2-n-heptyl-3-isothiazolone,

2-cyclopentyl-3-isothiazolone,

2-(4'-chlorophenyl)-3-isothiazolone,

2-(2',4'-dichlorophenyl)-3-isothiazolone,

2-(2',3'-dichlorophenyl)-3-isothiazolone,

2-(2',5'-dichlorophenyl)-3-isothiazolone,

2-(3'-chlorophenyl)-3-isothiazolone,

2-phenyl-3-isothiazolone,

2-(2'-chlorophenyl)-3-isothiazolone,

2-n-pentyl-3-isothiazolone,

2-i-propyl-3-isothiazolone,

2-(2-hydroxyethyl)-3-isothiazolone,

2-(2-bromoethyl)-3-isothiazolone,

2-(1,2,2,2-tetrachloroethyl)-3-isothiazolone,

2-chloromethyl-3-isothiazolone,

2-(2-dimethylaminoethyl)-3-isothiazolone,

4,5-dichloro-2-t-octyl-3-isothiazolone,

4-chloro-2-n-octyl-3-isothiazolone,

4-bromo-2-n-octyl-3-isothiazolone,

4-bromo-2-(4'-chlorophenyl)-3-isothiazolone,

4-bromo-2-t-butyl-3-isothiazolone,

2-(2,2,2-trichloro-1-hydroxyethyl)-3-isothiazolone,

2-(2,2,2-tribromo-1-hydroxyethyl)-3-isothiazolone,

2-trichlorobenzyl-3-isothiazolone,

2-sec-butyl-3-isothiazolone,

4-methyl-2-isopropyl-3-isothiazolone,

2-(4'-methylphenyl)-3-isothiazolone,

2-hydroxymethyl-3-isothiazolone, and

2-[2-(N,N-diethylamino)ethyl]-3-isothiazolone.

All of the isothiazolones with the exception of the hydroxyalkyl,alkenyl, and alkynyl derivatives, can be prepared by the cyclization ofa substituted disulfide-amide having the formula ##STR2## wherein H andH' are hydrogen or lower alkyl and Y' can be any of the groups embracedby Y in Formula I, as defined above, except alkenyl, alkynyl, and lowerhydroxyalkyl groups. The cyclization is accomplished by reacting thedisulfide-amide with a halogenating agent. Any halogenating agent may beemployed in this reaction. Typical halogenating agents include chlorine,bromine, sulfuryl chloride, sulfuryl bromide, N-chlorosuccinimide,N-bromosuccinimide, iodine monochloride, and the like. Bromine andchlorine are preferred halogenating agents. Cyclization of thedisulfide-amide will take place when 3 mole equivalents of halogenatingagent are employed in the reaction. By providing an excess ofhalogenating agent, the isothiazolone may be halogenated at the 4-and/or 5-positions of Formula I. Where 5 mole equivalents ofhalogenating agent are available, mono-halogenation can take place. Fordi-halogenation, 7 mole equivalents of halogenating agent are required.Preparation of isothiazolones having the 4- and 5-positions substitutedwith different halogens is achieved by the halogenation of anisothiazolone already halogenated at one of the two positions. Forexample, if a 4-bromo-5-chloro-3-isothiazolone is desired, it can beobtained by bromination of a 5-chloro-3-isothiazolone or thechlorination of a 4-bromo-3-isothiazolone. The starting 3-isothiazoloneis prepared by the cyclization of a disulfideamide as described above.The cyclization process will proceed over a broad temperature range andtemperature is not critical to the reaction. Generally, the cyclizationwill be carried out in the range of 0° to 100° C. The reaction iscarried out in an inert non-aqueous solvent, such as, for example,benzene, toluene, xylene, ethyl acetate, or ethylene dichloride. Inaddition, the 4,5-dihalo-3-isothiazolones may be prepared by in situhalogenation of a 5-halo-3-isothiazolone, as described for type Acompounds.

In addition, isothiazolones of the formula ##STR3## can be prepared bythe method of Goerdeler and Mittler as described in Chem. Ber., 96,944-954 (1963). In formula II, Y' can be any of the groups embraced by Yin Formula I, as defined above, except alkenyl, alkynyl, and lowerhydroxyalkyl groups, Z is hydrogen or lower alkyl and Z' is lower alkyl,

This preparation involves the halogenation of a substitutedβ-thioketoamide in an inert organic ester solvent, such as ethylacetate. The β-thioketoamide can be represented by the formula ##STR4##wherein Z, Z' and Y' are as defined above.

Another process for preparing some of the compounds of this invention isthat of Crow and Leonard as set forth in the Journal of OrganicChemistry, 30, 2660-2665 (1965). This method entails the converting of aβ-substituted thiocyanoacrylamide or thiosulfatoacrylamide to theisothiazolone. This reaction can be represented by the followingequation: ##STR5## wherein Y is as defined in Formula I, Z" is hydrogenor lower alkyl and M is --SCN or --S₂ O₃.sup.⊖. Generally, conversion ofthe substituted thiocyanoacrylamide or thiosulfatoacrylamide to theisothiazolone is achieved by the treating the amide with an acid, suchas for example, sulfuric acid. However, when H is SCN, transition metalsalts, such as nickel sulfate, ferrous sulfate, ferric sulfate, coppersulfate and the like, can also be readily utilized, and when H is S₂O₃.sup.⊖, iodine can be used. Preparation of the thiocyanoacrylamide andthiosulfatoacrylamido as taught by Crow and Leonard involves thereacting of a substituted propiolamide with thiocyanic acid orthiosulfate.

Some of the 2-hydroxyalkyl-3-isothiazolone derivatives disclosed hereinare obtained by reacting formaldehyde or an alkylene oxide, such asethylene oxide, propylene oxide and the like, with a3-hydroxyisothiazole. This reaction can be carried out in water,alcohols, or non-hydroxylic solvents, such as benzene, toluene, ethylacetate, ether, and hexane, and preferably a reaction temperature ofabout -20° C. to 100° C. is chosen. In order to facilitate the reaction,a basic catalyst, such as potassium carbonate, calcium carbonate,potassium hydroxide, or sodium hydroxide, can be advantageouslyemployed.

Preparation of the 2-haloalkyl-3-isothiazolones of the invention can beaccomplished by reacting a 2-hydroxyalkyl-3-isothiazolone with a halogencompound such as thionyl chloride, phosphorus tribromide, phosphorustrichloride and the like. Either the halogen compound or anon-hydroxylic solvent can be used as a solvent for the reaction, whichwill generally be carried out at about -20° C. to 100° C.

Some of the 2-alkylamino and 2-arylamino-3-isothiazolones of theinvention can be obtained by reacting formaldehyde and an appropriateamine with a 3-hydroxyisothiazole. An equimolar ratio of the reactantsis generally used, and water, alcohols, or non-hydroxylic solvents canbe employed. The reaction proceeds smoothly over a temperature range ofabout -10° C. to 100° C.

The 2-carbamoxyalkyl-3-isothiazolones of the invention are obtained byreacting an isocyanate with a 2-hydroxyalkyl-3-isothiazolone. Althoughan equimolar ratio of reactants is generally used, an excess of theisocyanate can be used. Non-hydroxylic solvents, such as aromatic andaliphatic hydrocarbons and chlorinated hydrocarbons, ethers, esters,nitriles, and amides can be used in carrying out the reaction. Atemperature of about -20° C. to 80° C. is usually chosen for thereaction, and when higher molecular weight or more complex isocyanatesare used as reactants, a catalyst, such as a tertiary amine, can beadvantageously employed to facilitate reaction.

The 2-alkenyl and 2-alkynyl-3-isothiazolones of the invention can beprepared by reacting an appropriate haloalkene or haloalkyne with a3-hydroxyisothiazole. The reaction can be carried out in water,alcohols, and non-hydroxylic solvents and a temperature of about -10° C.to 100° C. is generally utilized. An acid-accepting base, such as ametal hydride, a metal hydroxide, or a tertiary amine, is also generallyused to catalyze the reaction.

The 2-vinyl-3-isothiazolones of the invention are prepared by reacting avinyl acetate with a 3-hydroxyisothiazole. The vinyl acetate or othernon-hydroxylic compounds can be used as a solvent for the reaction whichcan be carried out at a temperature of about -10° C. to 100° C. Mercuricsalts can be advantageously used to catalyze the reaction.

Various 3-isothiazolone derivatives can be obtained by the reactiveaddition of a vinyl compound having the formula CH₂ ═CH--C with a3-hydroxyisothiazole. Q can represent groups such as N-pyrrolidonyl,N-isothiazolonyl, and the like. The addition is generally carried out ata temperature of about -20° C. to 100° C. using water, alcohols, ornon-hydroxylic compounds as solvents. For sluggish reactants, a basiccatalyst, such as a tertiary amine or a quaternary hydroxide can be usedto aid the reaction.

The 2-(2,2,2-trihalo-1-hydroxyalkyl)-3-isothiazolones of the inventionare prepared by reacting a trihaloacetaldehyde with a3-hydroxyisothiazole, in water, alcohols, or non-hydroxylic compounds assolvents, at a temperature of about -10° C. to 100° C. Theseisothiazolones can be dehydrated to form the corresponding unsaturated2-(1,2,2-trihaloalkenyl)-3-isothiazolones by reacting them with athionyl halide.

By way of demonstration, the following examples are offered toillustrate this invention and are not to be construed as limitationsthereof. In Table I, the compounds prepared by the above-describedprocesses and constituting Examples 1 through 103 are named and theirembodiments indicated. Table II lists the elemental analyses and meltingpoints (or boiling ranges) for the examples previously described inTable I. Specific illustrative preparations of Examples 7, 9, 11, 15,16, 37, 38, 52, 57, 60, 62, 67, and 98 are set forth below.

EXAMPLE 7 Preparation of 5-chloro-2-methyl-3-isothiazolone

To an ethylene dichloride (1 liter) slurry ofdithio-N,N'-dimethyldipropionamide, 70.9 g. (0.3 mole), there was addedat 10°-15° C. over 1.5 hour 121.5 g. (0.9 mole) of sulfuryl chloride.After addition, the reaction slurry was allowed to warm to 20°-25° C.and stirred overnight to assure completion of the reaction. The slurrywas then filtered to give 37.1 g. of 2-methyl-3-isothiazolonehydrochloride. The ethylenedichloride filtrate, upon evaporation toapproximately one-half volume, yielded an additional quantity (30.5 g.)of less pure hydrochloride. Complete evaporation of the ethylenedichloride filtrate gave 24.7 g. of oily residue which upon sublimationat 0.1 mm. (40°-60° C.) gave 11.5 g. of5-chloro-2-methyl-3-isothiazolone, m.p. 44°-47° C.

EXAMPLE 9 Preparation of 4,5-dichloro-2-methyl-3-isothiazolone

To a solution of 5.2 g. (0.045 mole) of 2-methyl-3-isothiazolone in 100ml. of ethyl acetate at -70° C. was added 6.08 g. (0.045 mole) ofsulfuryl chloride over 45 min. The reaction was stirred at thistemperature for 4 hours, then was allowed to come to room temperature.The ethyl acetate was removed by evaporation from the resulting slurry,and the solid thus obtained was extracted with benzene. Filtration andevaporation of the benzene gave 4 g. (96% based upon sulfuryl chloride)of 4,5-dichloro-2-methyl-3-isothiazolone, which upon crystallizationfrom ligroin (90°-120° ) gave pure material, m.p. 114°-17°.

EXAMPLE 11 Preparation of4-methyl-2-(3,4-dichlorophenyl)-3-isothiazolone

Dithio-(3',4'-dichloro)-diisobutyranilide, 12.8 g. (0.0234 mole), wasslurried in 250 ml. of ethylene dichloride, and a solution of 9.5 g.(0.0702 mole) of sulfuryl chloride in 25 ml. of ethylene dichloride wasadded dropwise at 25°-30° C. over one hour. The slurry was stirredovernight to insure completion of the reaction and then filtered to give8.6 g. of gray 2-(3,4-dichlorophenyl)-4-methyl-3-isothiazolone, m.p.160°-161° C. The product was crystallized from ethyl acetate to give am.p. of 161°-163° C.

EXAMPLE 15 Preparation of 4-bromo-2-methyl-3-isothiazolone

To a solution of 5 g. (0.043 mole) of 2-methyl-3-isothiazolone in 100ml. of ethylene dichloride at 0°-5° was added dropwise over a period of35 min. a solution of 6.94 g. (0.043 mole) of bromine in 25 ml. ofethylene dichloride. When the addition was complete the mixture wasallowed to warm to room temperature and stir overnight. The mixture wasfiltered giving 1.6 g. (14%) of 4-bromo-2-methyl-3-isothiazolonehydrobromide as red solid. The hydrobromide was dissolved in water andextracted continuously with ether. Evaporation of the extract gave 1.3g. of 4-bromo-2-methyl-3-isothiazolone, m.p. 94-97 from ligroin(90°-120°). NMR analysis of this bromo compound confirmed its structureas the 4-bromo isomer by the 5-H absorption at 1.87↑ and showed noabsorption for a 4 proton.

EXAMPLE 16 Preparation of 2-hydroxymethyl-3-isothiazolone

Aqueous formaldehyde (37 percent), 4.5 g. (0.056 mole) was diluted with25 ml. of water, and then 3.8 g. (0.023 mole) of potassium carbonate wasadded. Thereafter, 5.5 g. (0.055 mole) of 3-hydroxyisothiazole was addedto the formaldehyde solution in one portion. A solid precipitate soonformed, and after 2 hours stirring, was filtered off to yield 1.8 g. ofwhite solid 2-hydroxymethyl-3-isothiazolone.

EXAMPLE 37 Preparation of 4-chloro-2-t-octyl-3-isothiazolone

To a solution of 10.7 g. (0.05 mole) of 2-t-octyl-3-isothiazolone in 100ml. of chloroform was added in a single portion 13.3 g. (0.1 mole) ofN-chlorosuccinimide. The mixture warmed somewhat but did not requirecooling. After stirring for 2.5 hours the mixture was filtered giving7.5 g. of crude succinimide. The filtrate was evaporated under reducedpressure and the residue dissolved in ether. The ether solution wasextracted with water, dried over anhydrous magnesium sulfate, andevaporated to a mixture of solid and oil. The oil was washed from thesolid with hexane leaving 1.7 g. (14%) of white4-chloro-2-t-octyl-3-isothiazolone, m.p. 137-140.

NMR analysis confirmed the 4-chloro structure of4-chloro-2-t-octyl-3-isothiazolone by the 5-H absorption at 2.18↑.

EXAMPLE 33 Preparation of 4-bromo-2-t-octyl-3-isothiazolone

To a solution of 10.7 g. (0.05 mole) of 2-t-octyl-3-isothiazolone in 100ml. of chloroform was added in a single portion 17.8 g. (0.1 mole) ofN-bromosuccinimide, and a slight exotherm was observed. After stirringfor 3 hours the mixture was filtered, and the solid washed with ether.The chloroform filtrate and ether washings were then evaporated to givea total of 7.8 g. (54%) of 4-bromo-2-t-octyl-3-isothiazolone, m.p.133-142.

NMR analysis of this product showed 5-H absorption at 2.00↑, with noobservable 4-H present.

EXAMPLE 52 Preparation of2-(3',4'-dichloroanilinomethyl)-3-isothiazolone

A 5.0 gm. (0.05 mole) sample of 3-isothiazolone and 8.3 gm. (0.05 mole)of 3,5-dichloroaniline were dissolved in 7.0 ml. methanol. The solutionwas cooled to 10° C. and 0.05 mole of aqueous formaldehyde solution wasslowly added. The mixture was cooled to 0° C., and the precipitatedwhite solid was filtered off to give 8.9 g. (65%) of2-(3',4'-dichloroanilinomethyl)-3-isothiazolone, m.p. 120°-2° C.

EXAMPLE 57 Preparation of 2-N-phenylcarbamoxymethyl-3-isothiazolone

In 50 ml. of benzene was mixed 5.25 g. (0.04 mole) of2-hydroxymethyl-3-isothiazolone and 4.75 g. (0.04 mole) ofphenylisocyanate. A few drops of triethylamine were added, and themixture was heated at 50° C. for five hours. The white precipitate wasthen filtered off, washed with benzene, and dried to give 3.3 g. (30%)of 2-H-phenylcarbamoxymethyl-3-isothiazolone, m.p. 189°-90° from ethylacetate.

EXAMPLE 60 Preparation of 2-allyl-3-isothiazolone

To a solution of 20.2 g. (0.20 mole) of 3-hydroxyisothiazole in 100 ml.of methanol was added at 25° C. 47.5 g. (0.22 mole) of sodium methoxide(25% in methanol). To the resulting solution was then added dropwise24.2 g. (0.20 mole) of allyl bromide in 50 ml. of methanol. Afterstirring for several hours, the methanol was removed by distillation,and the residue was taken up in ether. After washing with water, theether was dried and removed under vacuum. The crude oil residue was thendistilled to give 2.8 g. (10%) of 2-allyl-3-isothiazolone, b.p. 75° C.(0.25 mm).

EXAMPLE 62 Preparation of 2-vinyl-3-isothiazolone

In 300 ml. of vinyl acetate was dissolved 5.0 g. of mercuric acetate bygentle refluxing. The solution was then cooled to 0° C. and 0.4 ml. of30% fuming sulfuric acid was added, followed by 20 g. (0.20 mole) of3-hydroxyisothiazole. During five days the solution was heated at 50° C.The solution was then cooled and 4.5 g. of sodium acetate was added.After stirring for three hours, the mixture was filtered, and the excessvinyl acetate was removed under vacuum. Distillation of the residue gave11.3 g. (45%) of 2-vinyl-3-isothiazolone, b.p. 90-94 (0.05 mm). Thedistilled product solidified and after crystallization from ether-hexanehad m.p. 56°-58° C.

EXAMPLE 67 Preparation of2-{1-[1-(2-pyrrolidononyl)]ethyl}-3-isothiazolone

To a solution of 10.1 g. (0.10 mole) of 3-hydroxyisothiazole in 50 ml.of benzene was added 11.1 g. (0.10 mole) of 1-vinyl-pyrrolidinone. Aslight exotherm was observed and after stirring at 25° C. for 12 hours,a white precipitate had formed. After filtration and drying, 11.5 g.(54%) of 2-{1-[1-(2-pyrrolidinonyl)]ethyl}-3-isothiazolone, m.p. 85°-88°C., was obtained.

EXAMPLE 98 Preparation of2-(1-hydroxy-2,2,2-trichloroethyl)-3-isothiazolone

To a solution of 5.0 g. (0.05 mole) of 3-hydroxyisothiazole in 50 ml. ofbenzene was added at 25° C. over 0.5 hour 7.8 g. (0.05 mole) of chloralin 15 ml. of benzene. A white precipitate formed, and after two hoursstirring at 25° C., the reaction was filtered to give 11.5 g. (90%) of2-(1-hydroxy-2,2,2-trichloroethyl)-3-isothiazolone, m.p. 117°-18° C.from ether-hexane.

                                      TABLE I                                     __________________________________________________________________________    3-ISOTHIAZOLONE EXAMPLES                                                       ##STR6##                                                                     Example                                                                       No.  R  R'                                                                                Y              Name                                               __________________________________________________________________________    1    H  H C.sub.3 H.sub.7 -n                                                                           2-n-propyl-3-isothiazolone                           2    H  H C.sub.4 H.sub.9 -t                                                                           2-t-butyl-3-isothiazolone                            3    H  H C.sub.4 H.sub.9 -n                                                                           2-n-butyl-3-isothiazolone                            4    H  H C.sub.6 H.sub.11                                                                             2-cyclohexyl-3-isothiazolone                         5    H  H C.sub.8 H.sub.17 -t                                                                          2-t-octyl-3-isothiazolone                            6    H  H CH.sub.2 C.sub.6 H.sub.4                                                                     2-benzyl-3-isothiazolone                             7    H  Cl                                                                              CH.sub.3       5-chloro-2-methyl-3-isothiazolone                    8    H  Cl                                                                              CH.sub.2 C.sub.6 H.sub.4                                                                     5-chloro-2-benzyl-3-isothiazolone                    9    Cl Cl                                                                              CH.sub.3       4,5-dichloro-2-methyl-3-isothiazolone                10   CH.sub.3                                                                         H CH.sub.3       2,4-dimethyl-3-isothiazolone                         11   CH.sub.3                                                                         H C.sub.6 H.sub.3 Cl.sub.2 -3,4                                                                4-methyl-2-(3,4-dichlorophenyl)-3-isothiazolone      12   H  H C.sub.6 H.sub.3 Cl.sub.2 -3,4                                                                2-(3,4-dichlorophenyl)-3-isothiazolone               13   Cl Cl                                                                               CH.sub.2 C.sub.6 H.sub.5                                                                    4,5-dichloro-2-benzyl-3-isothiazolone                14   Br Cl                                                                              CH.sub.3       4-bromo-5-chloro-2-methyl-3-isothiazolone            15   Br H CH.sub.3       4-bromo-2-methyl-3-isothiazolone                     16   H  H CH.sub.2 OH    2-hydroxymethyl-3-isothiazolone                      17   H  H CH.sub.2 CH.sub.2 N(CH.sub.2 CH.sub.3).sub.2                                                 2-(β-diethylaminoethyl)-3-isothiazolone         18   H  H C.sub.3 H.sub.7 . HCl                                                                        2-n-propyl-3-isothiazolone hydrochloride             19   H  Cl                                                                              CH.sub.3 . HCl 5-chloro-2-methyl-3-isothiazolone hydrochloride      20   H  H C.sub.2 H.sub.5 . HCl                                                                        2-ethyl-3-isothiazolone hydrochloride                21   H  H CH.sub.3 . HCl 2-methyl-3-isothiazolone hydrochloride               22   H  H CH.sub.2 C.sub.6 H.sub.5 . HCl                                                               2-benzyl-3-isothiazolone hydrochloride               23   H  H C.sub.12 H.sub.25 -n                                                                         2-n-dodecyl-3-isothiazolone                          24   H  H C.sub.14 H.sub.29 -n                                                                         2-n-tetradecyl-3-isothiazolone                       25   H  H CH.sub.2C.sub.6 H.sub.4Cl-4                                                                  2-(4-chlorobenzyl)-3-isothiazolone                   26   H  H CH.sub.2C.sub.6 H.sub.4Cl-2                                                                  2-(2-chlorobenzyl)-3-isothiazolone                   27   H  H CH.sub.2C.sub.6 H.sub.3Cl.sub.2 -2,4                                                         2-(2,4-dichlorobenzyl)-3-isothiazolone               28   H  H CH.sub.2C.sub.6 H.sub.3Cl.sub.2 3,4                                                          2-(3,4-dichlorobenzyl)-3-isothiazolone               29   H  H CH.sub.2C.sub.6 H.sub.4OCH.sub.3 -4                                                          2-(4-methoxybenzyl)-3-isothiazolone                  30   H  H CH.sub.2 C.sub.6 H.sub.4CH.sub.3 -4                                                          2-(4-methylbenzyl)-3-isothiazolone                   31   H  H CH.sub.2CH(C.sub.4 H.sub.9)C.sub.2 H.sub.5                                                   2-(2-ethylhexyl)-3-isothiazolone                     32   H  H CH.sub.2 CH.sub.2C.sub.6 H.sub.5                                                             2-(2-phenylethyl)-3-isothiazolone                    33   H  Cl                                                                              CH.sub.2 CH.sub.2C.sub.6 H.sub.5                                                             2-(2-phenylethyl)-5-chloro-3-isothiazolone           34   H  H CH(CH.sub.3)C.sub.6 H.sub.5                                                                  2-(1-phenylethyl)-3-isothiazolone                    35   H  H C.sub.10 H.sub.21 -n                                                                         2-n-decyl-3-isothiazolone                            36   H  H C.sub.8 H.sub.17 -n                                                                          2-n-octyl-3-isothiazolone                            37   Cl H C.sub.8 H.sub.17 -t                                                                          2-t-octyl-4-chloro-3-isothiazolone                   38   Br H C.sub.8 H.sub.17 -t                                                                          2-t-octyl-4-bromo-3-isothiazolone                    39   H  H C.sub.9 H.sub.19 -n                                                                          2-n-nonyl-3-isothiazolone                            40   H  Cl                                                                              C.sub.8 H.sub.17 -n                                                                          2-n-octyl-5-chloro-3-isothiazolone                   41   H  H C.sub.6 H.sub.4 NO.sub.2 4                                                                   2-(4-nitrophenyl)-3-isothiazolone                    42   H  H C.sub.6 H.sub.4 COOC.sub.2 H.sub.5 -4                                                        2-(4-carbethoxyphenyl)-3-isothiazolone               43   H  H CH.sub.2 CH.sub.2 CN                                                                         2-(2-cyanoethyl)-3-isothiazolone                     44   H  H CH.sub.2 CH.sub.2 CO.sub.2 CH.sub.3                                                          2-(2-carbomethoxyethyl)-3-isothiazolone              45   H  H CH.sub.2 CH.sub.2 CO.sub.2 CH.sub.3 . HCl                                                    2-(2-carbomethoxyethyl)-3-isothiazolone                                       hydrochloride                                        46   H  H CH.sub.2 CO.sub.2 CH.sub.3 . HCl                                                             2-carbomethoxymethyl-3-isothiazolone                                          hydrochloride                                        47   H  H CH.sub.2 CH.sub.2 OCH.sub.2 CH.sub.3 . HCl                                                   2-(2-ethoxyethyl)-3-isothiazolone hydrochloride      48   H  H CH.sub.2 CH.sub.2 OCH.sub.2 CH.sub.3                                                         2-(2-ethoxyethyl)-3-isothiazolone                    49   H  H                                                                                ##STR7##      2-(3',3',5'-trimethylcyclohexyl)-3-isothiazolone                              3                                                    50   H  H CH.sub.2 CH.sub.2 OC.sub.6 H.sub.5                                                           2-(2-phenoxyethyl)-3-isothiazolone                   51   H  H CH.sub.2 CH.sub.2 OCH.sub.3                                                                  2-(2-methoxyethyl)-3-isothiazolone                   52   H  H CH.sub.2 NHC.sub.6 H.sub.3Cl.sub.2 -3,4                                                      2-(3',4'-dichloroanilinomethyl)-3-isothiazolone      53   H  H CH.sub.2 NHC.sub.6 H.sub.4Cl-4                                                               2-(4'-chloroanilinomethyl)-3-isothiazolone           54   H  H CH.sub.2 NHC.sub.6 H.sub.4NO.sub.2 -4                                                        2-(4'-nitroanilinomethyl)-3-isothiazolone            55   H  H                                                                                ##STR8##      2-morpholinomethyl-3-isothiazolone                   56   H  H                                                                                ##STR9##      2-piperidinomethyl-3-isothiazolone                   57   H  H CH.sub.2 OCONHC.sub.6 H.sub.5                                                                2-phenylcarbanoxymethyl-3-isothiazolone              58   H  H CH.sub.2 OCONHC.sub.6 H.sub.4Cl-3                                                            2-(3'-chlorophenylcarbanoxymethyl)-3-isothiazolon                             e                                                    59   H  H CH.sub.2 OCONHC.sub.6 H.sub.3Cl.sub.2 -3,4                                                   2-(3' ,4'-dichlorophenylcarbanoxymethyl)-3-isothi                             azolone                                              60   H  H CH.sub.2 CHCH.sub.2                                                                          2-allyl-3-isothiazolone                              61   H  H CH.sub.2 CCH   2-propynyl-3-isothiazolone                           62   H  H CHCH.sub.2     2-vinyl-3-isothiazolone                              63   H  Cl                                                                              CHCH.sub.2     5-chloro-2-vinyl-3-isothiazolone                     64   H  H CH.sub.2 OCH.sub.3                                                                           2-methoxymethyl-3-isothiazolone                      65   H  H CH.sub.2 CH.sub.2 CO.sub.2 H . HCl                                                           2-(2-carboxyethyl)-3-isothiazolone                                            hydrochloride                                        66   H  H CH.sub.2 CH.sub.2 CO.sub.2 C.sub.4 H.sub.9 -n                                                2-(2-carb-n-butoxyethyl-3-isothiazolone              67   H  H                                                                                ##STR10##     2-[1-N-pyrrolidonyl)ethyl]-3-isothiazolone           68   H  H                                                                                ##STR11##     2-[1-(N-isothiazolonyl)ethyl]-3-isothiazolone        69   H  H CClCCl.sub.2   2-(1,2,2-trichlorovinyl)-3-isothiazolone             70   H  H CHBrCH.sub.2 OCH.sub.3                                                                       2-(1-bromo-2-methoxyethyl)-3-isothiazolone           71   H  H CH.sub.2 CH.sub.2 Cl                                                                         2-(2-chloroethyl)-3-isothiazolone                    72   H  H CH.sub.2 CH.sub.2 Cl . HCl                                                                   2-(2-chloroethyl)-3-isothiazolone hydrochloride      73   H  H CH.sub.2 CH.sub.2 CH.sub.2 Cl                                                                2-(3-chloropropyl)-3-isothiazolone                   74   H  H                                                                                ##STR12##     2-cyclopropyl-3-isothiazolone                        75   H  H CH.sub.2 CH.sub.2 C.sub.6 H.sub.5Cl-4                                                        2-[2-(4'-chlorophenyl)ethyl]-3-isothiazolone         76   H  H C.sub.6 H.sub.13 -n                                                                          2-n-hexyl-3-isothiazolone                            77   H  H C.sub.7 H.sub.14 -n                                                                          2-n-heptyl-3-isothiazolone                           78   H  H                                                                                ##STR13##     2-cyclopentyl-3-isothiazolone                        79   H  H C.sub.6 H.sub.4Cl-4                                                                          2-(4'-chlorophenyl)-3-isothiazolone                  80   H  H C.sub.6 H.sub.3Cl.sub.2 -2,4                                                                 2-(2',4'-dichlorophenyl)-3-isothiazolone             81   H  H C.sub.6 H.sub.3Cl.sub.2 -2,3                                                                 2-(2',3'-dichlorophenyl)-3-isothiazolone             82   H  H C.sub.6 H.sub.3Cl.sub.2 -2,5                                                                 2-(2',5'-dichlorophenyl)-3-isothiazolone             83   H  H C.sub.6 H.sub.4Cl-3                                                                          2-(3'-chlorophenyl)-3-isothiazolone                  84   H  H C.sub.6 H.sub.5                                                                              2-phenyl-3-isothiazolone                             85   H  H C.sub.6 H.sub.4Cl-2                                                                          2-(2'-chlorophenyl)-3-isothiazolone                  86   H  H C.sub.5 H.sub.11 -n                                                                          2-n-pentyl-3-isothiazolone                           87   H  H C.sub.3 H.sub.7 -iso                                                                         2-i-propyl-3-isothiazolone                           88   H  H CH.sub.2 CH.sub.2 OH                                                                         2-(2-hydroxyethyl)-3-isothiazolone                   89   H  H CH.sub.2 CH.sub.2 Br . HBr                                                                   2-(2-bromoethyl)-3-isothiazolone hydrobromide        90   H  H CHClCCl.sub.3  2-(1,2,2,2-tetrachloroethyl)-3-isothiazolone         91   H  H CH.sub.2 Cl    2-chloromethyl-3-isothiazolone                       92   H  H CH.sub.2 N(CH.sub.3).sub.2                                                                   2-(2-dimethylaminoethyl)-3-isothiazolone             93   Cl Cl                                                                              C.sub.8 H.sub.17 -t                                                                          4,5-dichloro-2-t-octyl-3-isothiazolone               94   Cl H C.sub.8 H.sub.17 -n                                                                          4-chloro-2-n-octyl-3-isothiazolone                   95   Br H C.sub.8 H.sub.17 -n                                                                          4-bromo-2-n-octyl-3-isothiazolone                    96   Br H C.sub.6 H.sub.4Cl-4                                                                          4-bromo-2-(4'-chlorophenyl)-3-isothiazolone          97   Br H C.sub.4 H.sub.9 -t                                                                           4-bromo-2-t-butyl-3-isothiazolone                    98   H  H CHOHCCl.sub.3  2-(2,2,2-trichloro-1-hydroxyethyl)-3-isothiazolon                             e                                                    99   H  H CHOHCBr.sub.3  2-(2,2,2-tribromo-1-hydroxyethyl)-3-isothiazolone    100  H  H CH.sub.2 C.sub.6 H.sub.2 Cl.sub.3                                                            2-trichlorobenzyl-3-isothiazolone                    101  H  H CH.sub.2 C.sub.6 H.sub.4OCH.sub.3 -4 . HCl                                                   2-(4'-methoxybenzyl)-3-isothiazolone                                          hydrochloride                                        102  H  H C.sub.4 H.sub.9 sec                                                                          2-sec-butyl-3-isothiazolone                          103  CH.sub.3                                                                         H C.sub.3 H.sub.7 -iso                                                                         4-methyl-2-iso-propyl-3-isothiazolone                104  H  Cl                                                                              C.sub.8 H.sub.17 -n                                                                          5-chloro-2-n-octyl-3-isothiazolone                   105  Cl Cl                                                                              C.sub.6 H.sub.11                                                                             4,5-dichloro-2-cyclohexyl-3-isothiazolone            106  CH.sub.3                                                                         Cl                                                                              C.sub.6 H.sub.4 Cl-3                                                                         5-chloro-2-(3-chlorophenyl)-4-methyl-3-isothiazol                             one                                                  __________________________________________________________________________

                                      TABLE II                                    __________________________________________________________________________     Example                                                                           Melting                                                                              ANALYSIS.sup.1                                                    No.  Point in ° C.                                                                   C     H     N     S   Cl or Br                                  __________________________________________________________________________    1    65-68  50.28(50.3)                                                                         6.17(6.33)                                                                          9.83(9.78)                                                                          22.30(22.3)                                     2    75-79  46.76(53.5)                                                                         6.76(7.06)                                                                          8.42(8.92)                                                                          16.43(20.4)                                     3    99     52.10(53.5)                                                                         7.11(7.06)                                                                          8.54(8.92)                                                                          19.35(20.4)                                     4    109-111                                                                              58.96(59.0)                                                                         7.20(7.10)                                                                          7.59(7.65)                                                                          17.41(17.5)                                     5    97-100 61.91(62.0)                                                                         9.03(9.0)                                                                           6.55(6.57)                                                                          14.89(15.03)                                    6    78-80  62.82(62.8)                                                                         4.84(4.7)                                                                           7.22(7.3)                                                                           16.97(16.75)                                    7    44-47  32.80(32.1)                                                                         2.64(2.7)                                                                           9.55(9.5)                                                                           20.79(21.4)                                                                         23.48(23.7)                               8    57-59  53.19(53.2)                                                                         3.46(3.6)                                                                           6.18(6.2)                                                                           13.80(14.2)                                                                         15.01(15.74)                              9    114-117                                                                              27.31(26.1)                                                                         1.43(1.6)                                                                           7.22(7.6)                                                                           17.42(17.4)                                                                         37.89(38.6)                               10   58-68  45.99(46.4)                                                                         5.77(5.4)                                                                           11.38(10.8)                                                                         24.39(24.8)                                     11   161-163                                                                              45.92(45.8)                                                                         2.88(2.7)                                                                           5.59(5.35)                                                                          11.91(12.2)                                                                         26.93(27.1)                               12   131-133                                                                              43.95(43.9)                                                                         1.95(2.05)                                                                          5.66(5.69)                                                                          13.09(13.0)                                     13   62-64  47.10(46.1)                                                                         2.86(2.7)                                                                           5.35(5.4)                                                                           12.48(12.3)                                                                         26.40(27.4)                               14   86-87  22.24(21.0)                                                                         1.81(1.3)                                                                           5.82(6.1)                                                                           13.83(14.0)                                                                         15.63(15.5)                                                                   **32.78(34.9)                             15   94-97  25.68(24.7)                                                                         1.75(2.0)                                                                           7.22(7.2)   **39.78(41.2)                             16   124-126                                                                              36.43(36.5)                                                                         3.75(3.7)                                                                           10.44(10.7)                                                                         24.30(24.4)                                     17   oil    52.70(54.1)                                                                         6.35(8.06)                                                                          14.17(14.0)                                                                         13.70(16.0)                                                                         N.E..sup.a 216(200)                       18   87-90  38.64(40.2)                                                                         5.88(5.61)                                                                          7.50(7.79)                                                                          16.85(17.83)                                                                        17.85(19.7)                               19   --     25.78(25.8)                                                                         3.17(2.7)                                                                           7.07(7.5)                                                                           14.9(17.2)                                                                          32.83(38.1)                               20   144-146                                                                              36.44(36.3)                                                                         4.80(4.8)                                                                           8.30(8.5)                                                                           19.20(19.3)                                                                         21.27(21.5)                               21   162-166                                                                              30.76(31.8)                                                                         4.27(4.0)                                                                           8.89(9.3)                                                                           20.79(21.2)                                                                         22.24(23.5)                               22   147-153                                                                              52.78(52.8)                                                                         4.60(4.39)                                                                          6.33(6.51)                                                                          14.21(14.1)                                                                         15.64(15.6)                               23   46-49  66.98(66.9)                                                                         9.90(10.0)                                                                          5.25(5.2)                                                                           11.72(11.90)                                    24   53-55  68.86(68.7)                                                                         10.61(10.5)                                                                         4.68(4.7)                                                                           10.88(10.8)                                     25   87-88  52.78(53.2)                                                                         3.54(3.6)                                                                           6.02(6.3)                                                                           14.59(14.2)                                                                         15.75(15.7)                               26   99-100 53.39(53.2)                                                                         3.69(3.6)                                                                           6.18(6.3)                                                                           13.95(14.2)                                                                         15.96(15.7)                               27   122-124                                                                              46.18(46.2)                                                                         2.80(2.7)                                                                           5.25(5.4)                                                                           12.46(12.3)                                                                         26.85(27.3)                               28   87-89  45.87(46.2)                                                                         2.62(2.7)                                                                           5.15(5.4)                                                                           12.12(12.3)                                                                         27.33(27.3)                               29   80-82  59.74(59.7)                                                                         4.96(5.0)                                                                           6.25(6.3)                                                                           14.58(14.5)                                     30   76-77  64.60(64.4)                                                                         5.32(5.4)                                                                           6.60(6.8)                                                                           15.60(15.6)                                     31   119-120                                                                              60.81(62.0)                                                                         8.88(8.9)                                                                           6.46(6.6)                                                                           14.70(15.0)                                          (.005mm)*                                                                32   76-78  64.35(64.5)                                                                         5.49(5.4)                                                                           6.75(6.8)                                                                           15.40(15.6)                                     33   55-59  55.22(55.2)                                                                         3.95(4.2)                                                                           5.62(5.9)                                                                           13.16(13.4)                                                                         14.51(14.8)                               34   138-140                                                                              62.52(64.5)                                                                         5.27(5.4)                                                                           6.45(6.8)                                                                           14.10(15.6)                                          (.003mm)*                                                                35   41-42  64.80(64.7)                                                                         9.31(9.6)                                                                           5.46(5.8)                                                                           13.49(13.2)                                     36   120(0.01mm)*                                                                         62.00(62.0)                                                                         8.81(8.9)                                                                           6.48(6.6)                                                                           14.89(15.0)                                     37   137-140                                                                              52.38(53.5)                                                                         7.81(7.3)                                                                           5.62(5.7)                                                                           --    12.19(14.5)                               38   138-142                                                                              44.76(45.2)                                                                         6.48(6.2)                                                                           4.58(4.8)                                                                           10.82(11.0)                                                                         **25.8(27.4)                              39   30-31  63.34(63.45)                                                                        9.40(9.29)                                                                          6.08(6.17)                                                                          13.97(14.10)                                    40   oil    55.05(53.4)                                                                         7.76(7.3)                                                                           5.76(5.7)                                                                           --    11.13(14.35)                              41   170-175 dec.                                                                         48.62(48.7)                                                                         2.78(2.72)                                                                          12.77(12.60)                                                                        --                                              42   141-142                                                                              58.12(57.8)                                                                         4.46(4.44)                                                                          5.70(5.62)                                                                          12.79(12.85)                                    43   85-6   46.68(46.8)                                                                         3.92(3.93)                                                                          18.25(18.2)                                                                         20.20(20.8)                                     44   125(0.01mm)*                                                                         44.69(44.9)                                                                         4.92(4.81)                                                                          7.41(7.49)                                                                          17.02(17.1)                                     45          37.81(37.6)                                                                         4.60(4.48)                                                                          6.21(6.26)                                                                          14.25(14.3)                                                                         14.64(15.9)                               46   90-93  34.39(34.4)                                                                         3.69(3.85)                                                                          6.81(6.68)                                                                          15.08(14.8)                                                                         16.30(16.9)                               47   99-103 40.11(39.9)                                                                         5.98(6.17)                                                                          6.62(6.65)                                                                          15.32(15.4)                                                                         15.37(16.9)                               48   90(0.04mm)*                                                                          48.95(48.8)                                                                         6.42(6.4)                                                                           7.98(8.1)                                                                           18.16(18.4)                                     49          63.61(64.0)                                                                         8.45(8.50)                                                                          6.12(6.22)                                                                          14.19(14.2)                                     50   78-81  59.61(59.7)                                                                         5.15(5.3)                                                                           6.23(6.3)                                                                           14.28(14.5)                                     51   89-112 44.82(45.3)                                                                         5.77(5.69)                                                                          8.66(8.79)                                                                          20.05(20.1)                                          (0.05mm)*                                                                52   120-122                                                                              43.75(43.6)                                                                         3.10(2.92)                                                                          10.18(10.2)                                                                         11.70(12.0)                                                                         25.69(25.7)                               53   93-94  49.74(49.7)                                                                         3.81(3.75)                                                                          11.59(11.6)                                                                         13.28(13.3)                                     54   196-197                                                                              47.74(47.7)                                                                         3.56(3.60)                                                                          16.66(16.8)                                                                         12.70(12.8)                                     55   114-115                                                                              48.39(48.0)                                                                         6.12(6.00)                                                                          13.90(14.0)                                                                         16.29(16.0)                                     56   95-98  55.00(54.5)                                                                         7.13(7.12)                                                                          14.35(14.2)                                                                         16.40(16.2)                                     57   189-190                                                                              52.95(52.8)                                                                         4.09(4.03)                                                                          11.09(11.2)                                                                         12.86(12.8)                                     58   168-170                                                                              46.26(46.4)                                                                         3.22(3.19)                                                                          9.74(9.84)                                                                          11.39(11.3)                                                                         12.49(12.4)                               59   196-199                                                                              41.42(41.4)                                                                         2.98(2.53)                                                                          8.78(8.77)                                                                          9.72(10.0)                                                                          22.22(22.0)                               60   95(0.5mm)*                                                                           50.12(51.2)                                                                         5.41(4.9)                                                                           9.87(9.9)                                                                           22.50(22.8)                                     61   110(0.2mm)*                                                                          50.14(51.7)                                                                         3.56(3.6)                                                                           10.27(10.1)                                                                         22.90(23.0)                                     62   56-58  47.41(47.2)                                                                         4.18(3.93)                                                                          10.99(11.1)                                                                         25.15(25.3)                                     63   84(0.02mm)*                                                                          37.80(37.2)                                                                         2.69(2.48)                                                                          7.85(8.65)                                                                          20.59(19.9)                                                                         21.95(20.1)                               64   80-9   41.14(41.1)                                                                         5.42(5.5)                                                                           9.70(9.60)                                                                          22.07(21.9)                                     65   156(dec)                                                                             34.11(34.4)                                                                         3.85(3.82)                                                                          6.59(6.68)                                                                          14.97(15.3)                                                                         16.98(16.9)                               66   150(0.01mm)*                                                                         52.50(52.4)                                                                         6.53(6.55)                                                                          6.05(6.11)                                                                          14.00(14.0)                                     67   91-92  51.07(51.0)                                                                         5.77(5.69)                                                                          13.25(13.2)                                                                         15.12(15.2)                                     68   130-133                                                                              40.45(42.2)                                                                         3.56(3.53)                                                                          11.75(12.3)                                                                         27.38(28.6)                                     69   91-93  25.93(26.1)                                                                         1.17(1.90)                                                                          6.10(6.05)                                                                          13.60(13.9)                                                                         48.8(45.4)                                70   104-106                                                                              22.99(22.6)                                                                         2.76(2.85)                                                                          4.28(4.38)                                                                          10.39(10.1)                                     71   100(0.02mm)*                                                                         36.63(36.7)                                                                         3.83(3.67)                                                                          8.48(8.56)                                                                          19.51(19.6)                                                                         21.36(21.7)                               72   132-135                                                                              29.77(30.0)                                                                         3.49(3.50)                                                                          6.72(7.00)                                                                          16.03(16.0)                                                                         35.14(35.5)                               73   45-48  40.07(40.6)                                                                         4.70(4.51)                                                                          7.72(7.83)                                                                          17.82(18.0)                                                                         21.11(20.0)                               74   65- 8  50.88(51.0)                                                                         4.78(4.99)                                                                          9.93(9.92)                                                                          22.92(22.7)                                     75          55.19(55.1)                                                                         4.23(4.21)                                                                          5.65(5.84)                                                                          13.35(13.4)                                                                         14.70(14.8)                               76   102(0.25                                                                             58.18(58.4)                                                                         7.62(8.11)                                                                          7.59(7.57)                                                                          17.02(17.3)                                          mm)*                                                                     77          60.53(60.3)                                                                         8.54(8.59)                                                                          7.07(7.02)                                                                          16.10(16.1)                                     78   118-19 56.84(56.8)                                                                         6.59(6.51)                                                                          8.32(8.28)                                                                          18.82(18.9)                                     79   142-44 51.02(51.1)                                                                         2.59(2.86)                                                                          6.43(6.63)                                                                          14.88(15.2)                                                                         16.55(16.8)                               80   155-57 44.20(43.9)                                                                         1.70(2.05)                                                                          5.52(5.69)                                                                          12.60(13.0)                                     81   130-34 43.70(43.9)                                                                         2.16(2.05)                                                                          5.60(5.69)                                                                          12.69(13.0)                                     82   145-47 43.71(43.9)                                                                         2.08(2.05)                                                                          5.58(5.69)                                                                          12.85(13.0)                                     83   119-22 51.26(51.1)                                                                         3.00(2.86)                                                                          6.55(6.63)                                                                          14.98(15.2)                                     84   19-2   61.31(61.1)                                                                         4.03(3.98)                                                                          7.86(7.90)                                                                          17.70(18.0)                                     85   90-1   51.06(51.1)                                                                         2.82(2.86)                                                                          6.55(6.63)                                                                          14.82(15.2)                                     86   118(0.01mm)*                                                                         56.09(56.14)                                                                        7.86(7.60)                                                                          8.14(8.19)                                                                          18.40(18.7)                                     87   84(0.15mm)*                                                                          50.46(50.3)                                                                         6.14(6.33)                                                  88   114-15 41.18(41.4)                                                                         4.91(4.83)                                                                          9.54(9.65)                                                                          21.80(21.2)                                     89          21.01(20.8)                                                                         2.69(2.42)                                                                          4.80(4.84)                                                                          11.31(11.0)                                                                         Br. 55.51(55.4)                           90   90-91  22.17(22.5)                                                                         1.35(1.12)                                                                          5.05(5.24)                                                                          11.91(12.0)                                                                         52.15(53.0)                               91   129-130                                                                              31.7(32.3)                                                                          2.87(2.68)                                                                          9.18(9.36)                                                                          21.4(20.8)                                                                          22.3(23.7)                                92   82-85  42.2(45.6)                                                                          5.34(6.33)                                                                          14.63(17.8)                                                                         21.3(20.2)                                      93          46.92(46.8)                                                                         6.36(6.03)                                                                          4.53(4.96)                                                                          10.52(11.3)                                                                         24.58(25.2)                               94          53.09(53.4)                                                                         7.75(7.33)  11.75(12.9)                                                                         11.74(14.3)                               95   42-5   45.57(45.3)                                                                         6.33(6.18)                                                                          4.44(4.79)                                                                          10.24(.0.9)                                                                         Br. 25.20(27.3)                           96   194-96 37.35(37.2)                                                                         1.60(1.73)                                                                          4.58(4.82)                                                                          10.98(11.0)                                                                         Br. 27.99(27.5)                           97   150-53(dec)                                                                          35.83(35.6)                                                                         4.27(4.27)                                                                          6.00(5.93)                                                                          13.28(13.56)                                                                        Br. 33.91(33.9)                           98   117-118                                                                              24.37(24.2)                                                                         1.90(1.61)                                                                          6.01(5.64)                                                                          12.58(12.9)                                                                         42.25(42.8)                               99          20.11(20.0)                                                                         1.60(1.47)                                                                          5.83(5.83)                                                                          11.35(13.3)                                                                         Br. 48.7(50.0)                            100  115-17 41.42(40.9)                                                                         1.71(2.00)                                                                          4.70(4.70)                                                                          11.13(10.8)                                                                         36.00(36.1)                               101  145-47 51.14(51.3)                                                                         4.51(4.66)                                                                          5.52(5.44)                                                                          12.08(12.4)                                                                         13.66(13.8)                               102  84.5(0.2mm)*                                                                         53.43(53.5)                                                                         7.13(7.05)                                                                          8.67(8.90)                                                                          19.42(20.4)                                     103  75-8(.09mm)*                                                                         50.94(53.5)                                                                         6.71(7.05)                                                                          8.42(8.90)                                                                          19.15(20.4)                                     __________________________________________________________________________     .sup.1 The number parenthesized represents the theoretical value, as          calculated, using the empirical formula of each compound.                     *Boiling point in ° C.                                                 **Analysis for bromine.                                                       *Neutralization equivalent.                                              

The novel isothiazolones and salts of this invention are biocidallyactive compounds, and as such, are suitable for the control of livingorganisms and particularly microorganisms. For this reason, they areespecially effective bactericidal, algaecidal, fungicidal, slimicidal,and pesticidal agents. Furthermore, these novel compounds possess theunexpected property of being resistant to inhibition by common additivesor contaminants, such as lecithin, normal horse serum, alkylbenzenesulfonates, water-soluble lanolin, sodium chromate, sodium nitrite,glycerol, propylene glycol and the like.

It was also determined that other isothiazolones, namely,2-methyl-3-isothiazolone and 2-ethyl-3-isothiazolone are likewiseeffective for the control of living organisms. These compounds weredescribed by Leonard and Crow in the Journal of Organic Chemistryarticle previously cited. However, Leonard and Crow did not attributeany biocidal properties to the compounds. The use of theseisothiazolones as biocides was unknown prior to the present invention.

Antibacterial and fungicidal activity were evaluated by the SerialDilution Test (Broth Titer Test) wherein a series of broths containingvarying dilutions of a test compound and an organism are halved startingwith 1:1000. The values obtained, which are also shown in Table III,represent the maximum dilution at which the compound under evaluationrenders complete control of the organism. Staphylococcus aureus (S.aureus) and Escherichia coli (E. coli) were the bacterial organismsemployed in this test, and the fungi employed were Aspergillus niger (A.niger), Rhizopus Stolonifer (Rhiz.), and Rhodetorula sp. (Rhode).

The 3-isothiazolones of the invention are also effective as algaecides.The algaecidal activity of the 3-isothiazolones was determined by theFitzgerald Test (Applied Microbiology, 7, 205-211, No. 4, 1959). TableIV shows the parts per million of the 3-isothiazolone under evaluationnecessary for complete control of the organism. Chlorella pyrenoidosa(Chlorella) and "Black" algae (Oscilatoria sp.) were the algae employedin this test.

                                      TABLE III                                   __________________________________________________________________________    MICROBIOLOGICAL ACTIVITY                                                                                BACTERIOSTATIC                                               FUNGISTATIC EVALUATION                                                                         EVALUATION                                          Example No.                                                                            A. niger Rhiz. (Rhodo.*)                                                                       S. aureus                                                                           E. coli                                       __________________________________________________________________________    1        1:16,000 1:32,000                                                                              1:8,000                                                                             1:32,000                                      2        1:8,000  1:8,000 1:4,000                                                                             1:8,000                                       3        1:16,000 1:125,000                                                                             1:8,000                                                                             1:32,000                                      4        1:8,000  1:64,000                                                                              1:32,000                                                                            1:8,000                                       5        1:16,000 1:8,000 1:8,000                                                                             1:1,000                                       6        1:4,000  1:64,000                                                                              1:32,000                                                                            1:64,000                                      7        1:125,000                                                                              1:500,000                                                                             1:128,000                                                                           1:256,000                                     8        1:32,000 1:125,000                                                                             1:64,000                                                                            1:64,000                                      9        1:16,000 1:16,000                                                                              1:32,000                                                                            1:64,000                                      10       1:2,000  1:4,000 1:2,000                                                                             1:4,000                                       11        --       --     1:256,000                                                                           1:1,000                                       12        --      1:8,000 1:512,000                                                                           1:2,000                                       13       1:16,000 1:250,000                                                                             1:128,000                                                                           1:64,000                                      14       1:32,000 >1:64,000                                                                             1:128,000                                                                           1:64,000                                      15       1:4,000  1:16,000                                                                               --    --                                           16       1:16,000 1:32,000                                                                              1:8,000                                                                             1:32,000                                      17        --       --     1:8,000                                                                             1:4,000                                       18       1:8,000  1:16,000*                                                                             1:32,000                                                                            1:8,000                                       20       1:4,000  1:8,000*                                                    21       1:64,000 1:64,000*                                                                             1:32,000                                                                            1:32,000                                      22       1:4,000  1:32,000*                                                                             1:32,000                                                                            1:32,000                                      23       1:16,000 1:250,000                                                                             1:1,000,000                                                                         1:1,000                                       24       1:2,000  1:8,000 1:1,000                                                                             <1:1,000                                      25       1:4,000  1:250,000                                                                             1:64,000                                                                            1:16,000                                      26       1:4,000  1:64,000                                                                              1:64,000                                                                            1:16,000                                      27       1:4,000  1:64,000                                                                              1:256,000                                                                           1:4,000                                       28       1:8,000  1:16,000                                                                              1:256,000                                                                           1:16,000                                      29       1:4,000  1:32,000                                                                              1:16,000                                                                            1:8,000                                       30       1:8,000  1:64,000                                                                              1:32,000                                                                            1:8,000                                       31       1:32,000 1:64,000                                                                              1:64,000                                                                            1:2,000                                       32       1:8,000  1:125,000                                                                             1:16,000                                                                            1:8,000                                       33       1:64,000 1:250,000                                                                             1:64,000                                                                            1:64,000                                      34       1:8,000  1:32,000                                                                              1:32,000                                                                            1:8,000                                       35       1:250,000                                                                              1:500,000                                                                             1:2,000,000                                                                         1:16,000                                      36       1:125,000                                                                              1:250,000                                                                             1:128,000                                                                           1:8,000                                       37       1:4,000  1:16,000                                                                              1:8,000                                                                             <1:1,000                                      38       1:4,000  1:8,000 1:8,000                                                                             <1:1,000                                      39       1:125,000                                                                              1:500,000                                                                             1:512,000                                                                           1:8,000                                       40       1:64,000 1:500,000                                                                             1:8,000                                                                             1:16,000                                      41        --       --     1:16,000                                                                            1:32,000                                      42        --       --     1:8,000                                                                             1:16,000                                      43       1:2,000  1:2,000 1:32,000                                                                            1:32,000                                      44       1:2,000  1:4,000 1:16,000                                                                            1:32,000                                      45       1:2,000  1:4,000 1:16,000                                                                            1:32,000                                      46       <1:2,000 1:2,000 1:16,000                                                                            1:32,000                                      48       1:4,000  1:8,000 1:8,000                                                                             1:32,000                                      49       1:32,000 1:125,000                                                                             1:32,000                                                                            1:1,000                                       50       1:8,000  1:32,000                                                                              1:32,000                                                                            1:8,000                                       51       1:1,000  1:2,000 1:16,000                                                                            1:16,000                                      52       1:16,000 1:32,000                                                                              1:8,000                                                                             1:8,000                                       53       1:16,000 1:16,000                                                                              1:8,000                                                                             1:8,000                                       54       1:16,000 1:16,000                                                                              1:8,000                                                                             1:8,000                                       55       1:8,000  1:16,000                                                                              1:8,000                                                                             1:16,000                                      56       1:16,000 1:32,000                                                                              1:8,000                                                                             1:16,000                                      57       1:8,000  1:8,000 1:8,000                                                                             1:8,000                                       58       1:8,000  1:16,000                                                                              1:4,000                                                                             1:2,000                                       59       1:4,000  1:16,000                                                                              1:512,000                                                                           1:1,000                                       60       1:8,000  1:32,000                                                                              1:16,000                                                                            1:16,000                                      61       1:4,000  1:32,000                                                                              1:32,000                                                                            1:32,000                                      62       1:8,000  1:64,000                                                                              1:125,000                                                                           1:125,000                                     63       1:125,000                                                                              1:125,000                                                                             1:125,000                                                                           1:250,000                                     64        --       --     1:32,000                                                                            1:32,000                                      65       <1:2,000 1:2,000 <1:1,000                                                                            1:1,000                                       66       1:8,000  1:16,000                                                                              1:32,000                                                                            1:8,000                                       67       1:8,000  1:16,000                                                                              1:8,000                                                                             1:16,000                                      68       1:4,000  1:8,000 1:32,000                                                                            1:16,000                                      69       1:4,000  1:8,000 1:2,000                                                                             1:4,000                                       70        --       --     1:16,000                                                                            1:16,000                                      71       1:8,000  1:16,000                                                                              1:64,000                                                                            1:64,000                                      72       1:4,000  1:8,000 1:32,000                                                                            1:32,000                                      73       1:8,000  1:16,000                                                                              1:32,000                                                                            1:16,000                                      74       1:4,000  1:8,000 1:32,000                                                                            1:32,000                                      75       1:16,000 1:250,000                                                                             1:64,000                                                                            1:1,000                                       76       1:64,000 1:125,000                                                                             1:32,000                                                                            1:8,000                                       77       1:250,000                                                                              1:250,000                                                                             1:64,000                                                                            1:8,000                                       78       1:16,000 1:32,000                                                                              1:16,000                                                                            1:32,000                                      79       1:2,000  1:16,000                                                                              1:64,000                                                                            1:8,000                                       80       <1:2,000 1:2,000 1:16,000                                                                            1:1,000                                       81       1:2,000  1:4,000 1:32,000                                                                            1:1,000                                       82       1:4,000  1:2,000 1:1,000                                                                             1:1,000                                       83       1:4,000  1:2,000 1:32,000                                                                            1:16,000                                      84       1:2,000  1:64,000                                                                              1:128,000                                                                           1:16,000                                      85       1:2,000  1:2,000 1:32,000                                                                            1:1,000                                       86       1:32,000 1:500,000                                                                             1:16,000                                                                            1:16,000                                      87       1:4,000  1:16,000                                                                               --    --                                           88       1:2,000  1:2,000 1:1,000                                                                             1:1,000                                       89       1:4,000  1:8,000 1:32,000                                                                            1:64,000                                      90       1:8,000  1:16,000                                                                              1:4,000                                                                             1:8,000                                       91       1:8,000  1:8,000 1:16,000                                                                            1:8,000                                       92       1:4,000  1:16,000                                                                              1:16,000                                                                            1:32,000                                      93       1:16,000  --     1:32,000                                                                            1:8,000                                       94       1:2,000  1:2,000 <1:1,000                                                                            1:1,000                                       95       1:125,000                                                                              1:64,000                                                                              1:256,000                                                                           1:16,000                                      96       1:4,000  1:4,000 1:32,000                                                                            1:2,000                                       97       1:8,000  1:32,000                                                                              1:32,000                                                                            1:8,000                                       98       1:8,000  1:16,000                                                                              1:4,000                                                                             1:16,000                                      99       1:8,000  1:16,000                                                                              1:8,000                                                                             1:16,000                                      100      1:8,000  1:32,000                                                                              1:256,000                                                                           <1:1,000                                      101      1:4,000  1:16,000                                                                              1:16,000                                                                            1:4,000                                       102      1:8,000  1:16,000                                                                              1:4,000                                                                             1:8,000                                       104      1:125,000                                                                              1:250,000                                                                             1:250,000                                                                           1:64,000                                      105      1:32,000.sup.1   1:32,000.sup.2                                      106      1:1,000,000.sup.1                                                                            1:64,000.sup.2                                        2-methyl-3-isothia-                                                           zolone   1:4,000  1:8,000 1:16,000                                                                            1:32,000                                      2-ethyl-3-isothia-                                                            zolone   1:4,000  1:16,000                                                                              1:8,000                                                                             1:16,000                                      __________________________________________________________________________      .sup.1 Mixed cultureA. niger, Rhiz.                                          .sup.2 Mixed cultureS. aureus, E. coli.                                  

                  TABLE IV                                                        ______________________________________                                        ALGA-STATIC ACTIVITY                                                          Example No.   Chlorella    Oscilatoria                                        ______________________________________                                         1            2            5                                                   3            5            5                                                   4            1            1                                                   5            10           --                                                  6            1            0.75                                                7            0.125        0.125                                               8            0.25         0.125                                               9            0.3          0.16                                               10            2            --                                                 11            2            1                                                  12            10           1                                                  13            0.25         0.125                                              14            0.25         0.125                                              15            0.5          < 0.125                                            16            5            5                                                  10            10           10                                                 20            5            --                                                 21            10           0.5                                                22            2            10                                                 23            2            < 0.5                                              24            2            < 0.5                                              25            1            1                                                  26            2            5                                                  27            5            2                                                  28            2            2                                                  29            10           10                                                 30            10           10                                                 31            5            10                                                 33            1            1                                                  34            --           10                                                 35            1            1                                                  36            1,5          2,5                                                38            1            1                                                  39            1            2                                                  40            1            2                                                  43            0.5          10                                                 44            --           10                                                 46            --           2                                                  48            --           0.5                                                49            2.5          10                                                 50            10           0.5                                                52            5            10                                                 53            5            5                                                  54            5            10                                                 55            2            2                                                  56            2            5                                                  57            2            < 0.5                                              58            2            < 0.5                                              59            10           < 0.5                                              60            1.25         2.5                                                62            5            0.3                                                63            < 0.5        <0.5                                               66            --           1                                                  67            5            --                                                 68            --           < 0.5                                              69            5            < 0.5                                              71            5            2                                                  73            10           5                                                  74            2.5          2.5                                                75            2.5          2.5                                                76            5            10                                                 77            2            0.5                                                78            5            10                                                 79            5            < 0.5                                              84            2            0.5                                                88            --           10                                                 89            --           5                                                  90            10           5                                                  91            10           --                                                 92            2            1                                                  93            5            2                                                  95            1            1                                                  96            10           0.5                                                97            5            2                                                  104           0.63         0.63                                               ______________________________________                                    

Further fungicidal activity on the part of the isothiazolones wasdemonstrated by their utilization in seed treatment applications. Byseed treatment is meant the disseminating of a biocidally activematerial over a seed subject to the attack of microorganisms, andparticularly fungi, in an amount which is effective to control suchmicroorganisms without deleteriously effecting the seed. In mostcircumstances, the biocidally active material, in this case, theisothiazolones or compositions thereof will be applied to the surfacearea of the seeds to be treated. This may be accomplished by varyingmeans common to the art, such as slurrying, soaking, dusting, sprayingand the like.

The amount of isothiazolone required so as to be effective in a seedtreatment application will vary depending upon condition, such as thetype of seed, the method of application, soil and atmospheric conditionsand the like. Generally, an application in the range of about 0.25 to 20ounces of active ingredient, namely, the isothiazolone per 100 pounds ofseed will be effective to control any undesirable microorganisms and toprovide protection to the seed. An application of active agent in therange of about 1.0 to 10 ounces per 100 pounds of seed is preferred.

An experimental technique was employed to demonstrate the effectivenessof these compounds for seed treatment. This involved slurrying two lotsof 100 corn seeds each with the isothiazolone under evaluation at alevel of 1 to 2 ounces per bushel of seed. For purposes of comparison,an untreated control was also included. After drying, the seeds wereplanted in a muck soil infested with Fythium and having a high moisturecontent. The corn seeds were maintained in this soil for a week at about10° C. so as to permit infection. Upon completion of this period oftime, the temperature was elevated and maintained at about 21° to 27° C.so as to permit germination of the non-infected seed. The number ofseeds germinating were counted after emergence of the plant and suchdata is set forth in Table V.

                  TABLE V                                                         ______________________________________                                        SEED TREATMENT                                                                                     No. of Plants                                                Test Compound    Germinating/200 Seeds                                    ______________________________________                                        untreated              10                                                     2-t-butyl-3-isothiazolone                                                                          100                                                      2-n-hexyl-3-isothiazolone                                                                          172                                                      2-n-octyl-3-isothiazolone                                                                          144                                                      2-t-octyl-3-isothiazolone                                                                          184                                                      2-n-nonyl-3-isothiazolone                                                                          164                                                      2-n-decyl-3-isothiazolone                                                                          153                                                      2-methyl-5-chloro-3-isothiazolone                                                                  150                                                      2-methyl-4,5-dichloro-3-isothiazolone                                                              140                                                      2-n-octyl-5-chloro-3-isothiazolone                                                                 150                                                      2-t-octyl-4-chloro-3-isothiazolone                                                                 160                                                      2-t-octyl-4-bromo-3-isothiazolone                                                                  148                                                      2-(2,4-dichlorobenzyl)-3-isothiazolone                                                               80                                                     2-(2-ethylhexyl)-3-isothiazolone                                                                   182                                                      2-(1-phenylethyl)-3-isothiazolone                                                                  176                                                      2-(2-phenylethyl)-3-isothiazolone                                                                  160                                                      2-cyclohexyl-3-isothiazolone                                                                       168                                                      2-benzyl-3-isothiazolone                                                                           136                                                      2-hydroxymethyl-3-isothiazolone                                                                    182                                                      ______________________________________                                    

Evaluation of the isothiazolones with regard to pesticidal activitydemonstrated their exceptional control of numerous nematodes, mites,insects, such as beetles and aphids, and the like. Some typicalnematodes, insects and mites which were controlled when contacted withthe compounds of this invention included the following: Northern rootknot nematode (Meloidogyae hapla), Mexican bean beetle (Epilachnavarivesta), black carpet beetle (Attagenus piceus), confused flourbeetle (Tribolium confusum), granary weevil (Sitophilus granarius), twospotted mite (Tetranychus urticae), house fly (Musca domestica),Southern armyworm (Prodenia eridania), German cockroach (Blattellagermanica), and green peach aphid (Myzus persicae).

Generally, control of a living organism is achieved in accordance withthis invention by contacting the organism with an isothiazolone in anamount which is effective to control said organism. Any of thetechniques known in the art can be employed to disseminate theisothiazolones in a manner so as to achieve the desired contact with theorganism to be controlled. Spraying and fumigating are typical of suchtechniques.

The compounds of this invention can be readily utilized as slimicides,algaecides, bactericides, and fungicides in any locus and particularlyin aqueous media, such as, for example, water-cooling systems, swimmingpools, paper pulp processes, aqueous polymer dispersions, water-basedpaints, and the like. In addition, these compounds and compositionscontaining them can function as, for example, fabric and leatherpreservatives, cosmetic preservatives, soap additives, sanitizingagents, such as in laundry soaps and detergents, and preservatives formetal working compounds, such as emulsifiable cutting oils,preservatives for fuels, fiber spin finish biocides, and the like.

In general, a locus subject to contamination by microorganisms can beprotected in accordance with this invention by incorporating into thelocus an isothiazolone or a salt of an isothiazolone in an amount whichis effective to control said microorganisms. The term "contamination" ismeant to include any attack by microorganisms which leads to a chemicalor physical breakdown or disintegration of the locus as well as theproliferation of the microorganisms within the locus without anaccompanying deleterious effect. The exact amount of isothiazolonerequired will, of course, vary with the medium being protected, themicroorganisms being controlled, the particular isothiazolones orcompositions containing the isothiazolones being employed and otherfactors. Typically, in a liquid medium, excellent control is obtainedwhen the isothiazolones are incorporated in the range of 0.1 to 10,000parts per million (ppm) or 0.00001 to 1% based on the weight of themedium. A range of 1 to 2000 ppm is preferred.

The term "control," as employed in the specification and claims of thisapplication is to be construed as the effect of any means whichadversely affects the existence or growth of any living organism ormicroorganism. This effect may comprise a complete killing action,eradication, arresting in growth, inhibition, reduction in number or anycombination thereof.

Outstanding fungistatic activity is exhibited by the isothiazolones whenthey were employed as paint preservatives and paint fungistats.Microbial activity in water-based and oil-based paint emulsions iseffectively inhibited when the isothiazolones are incorporated into thepaint. The isothiazolones are also highly active mildewcides for paintfilms when incorporated in paint formulations.

The isothiazolones of this invention are especially useful asagricultural fungicides. As such, they are particularly valuable whenformulated in a fungicidal composition. Such compositions normallycomprise an agronomically acceptable carrier and an isothiazolone ormixture of isothiazolones as the active agent. Where necessary ordesirable, surfactants or other additives may be incorporated to giveuniformly formulated mixtures. By "agronomically acceptable carrier" ismeant any substance which can be utilized to dissolve, dispense ordiffuse the chemical incorporated therein without impairing theeffectiveness of the toxic agent and which does no permanent damage tosuch environment as soil, equipment and agronomic crops.

For use as pesticides, the compounds of this invention are usually takenup in an agronomically acceptable carrier or formulated so as to renderthem suitable for subsequent dissemination. For example, theisothiazolones can be formulated as wettable powders, emulsionconcentrates, dusts, granular formulations, aerosols or flowableemulsifiable concentrates. In such formulations, the isothiazolones areextended with a liquid or solid carrier and, when desired, suitablesurfactants are likewise incorporated.

Compounds of this invention can be dissolved in a water-miscible liquid,such as ethanol, isopropanol, acetone, and the like. Such solutions areeasily extended with water.

The isothiazolones can be taken up on or mixed with a finely particledsolid carrier, as for example, clays, inorganic silicates, carbonates,and silicas. Organic carriers can also be employed. Dust concentratesare commonly made wherein isothiazolones are present in the range of 20to 80%. For ultimate applications, these concentrates are normallyextended with additional solid from about 1 to 20%.

Wettable powder formulations are made by incorporating the compounds ofthis invention in an inert, finely divided solid carrier along with asurfactant which may be one or more emulsifying, wetting, dispersing orspreading agents or blend of these. The isothiazolones are usuallypresent in the range of 10 to 80% by weight and the surfactants in from0.5 to 10% by weight. Commonly used emulsifying and wetting agentsinclude polyoxyethylated derivatives of alkylphenols, fatty alcohols,fatty acids, and alkylamines, alkylarene sulfonates and dialkylsulfosuccinates. Spreading agents include such materials as glycerolmannitan laurate and a condensate of polyglycerol and oleic acidmodified with phthalic anhydride. Dispersing agents include suchmaterials as the sodium salt of the copolymer of maleic anhydride and anolefin such as diisobutylene, sodium lignin sulfonate and sodiumformaldehydenaphthalene sulfonates.

One convenient method for preparing a solid formulation is to impregnatethe isothiazolone toxicant onto the solid carrier by means of a volatilesolvent, such as acetone. In this manner adjuvants, such as activators,adhesives, plant nutrients, synergists and various surfactants, can alsobe incorporated.

Emulsifiable concentrate formulations can be prepared by dissolving theisothiazolones of this invention in an agronomically acceptable organicsolvent and adding a solvent-soluble emulsifying agent. Suitablesolvents are usually water-immiscible and may be found in thehydrocarbon, chlorinated hydrocarbon, ketone, ester, alcohol and amideclasses of organic solvents. Mixtures of solvents are commonly employed.The surfactants useful as emulsifying agents may constitute about 0.5 to10% by weight of the emulsifiable concentrate and may be anionic,cationic or non-ionic in character. Anionic surfactants include alcoholsulfates or sulfonates, alkylarene sulfonates and sulfosuccinates.Cationic surfactants include fatty acid alkylamine salts and fatty acidalkyl quaternaries. Nonionic emulsifying agents include alkylene oxideadducts of alkylphenols, fatty alcohols, mercaptans and fatty acids. Theconcentration of the active ingredients may vary from 10 to 80%,preferably in the range of 25 to 50%.

For use as phytopathogenic agents, these compounds should be applied inan effective amount sufficient to exert the desired biocidal activity bytechniques well known in the art. Usually, this will involve theapplication of the isothiazolones to the locus to be protected in aneffective amount when incorporated in an agronomically acceptablecarrier. However, in certain situations it may be desirable andadvantageous to apply the compounds directly onto the locus to beprotected without the benefit of any substantial amount of carrier. Thisis a particularly effective method when the physical nature of theisothiazolone is such as to permit what is known as "low-volume"application, that is, when the compounds are in liquid form orsubstantially soluble in higher boiling solvents.

The application rate will, of course, vary depending upon the purposefor such application, the isothiazolones being utilized, the frequencyof dissemination and the like.

For use as agricultural bactericides, fungicides, nematocides, andinsecticides, dilute sprays can be applied at concentrations of 0.05 to20 pounds of the active isothiazolone ingredient per 100 gallons ofspray. They are usually applied at 0.1 to 10 pounds per 100 gallons andpreferably at 0.125 to 5 pounds per 100 gallons. In more concentratedsprays, the active ingredient is increased by a factor of 2 to 12. Withdilute sprays, applications are usually made to the plants until run-offis achieved, whereas with more concentrated or low-volume sprays thematerials are applied as mists.

The compounds of this invention may be utilized as the sole biocidalagents or they may be employed in conjunction with other fungicides,bactericides, algaecides, slimicides, insecticides, miticides, or withother comparable pesticides.

In field tests, the isothiazolones described above were effective incontrolling fungi which cause broccoli downy mildew (Peronosporaparasitica), bean powdery mildew (Prysiphe polygoni), wheat powderymildew (Erysiphe graminis), rice blast (Piricularia ovygae), and tomatoblights (Phytothora infestans) and (Alternaria solsni).

The following examples A through B illustrate several specificembodiments of the many uses of the isothiazolones of the invention.

EXAMPLE A Preservatives for Cutting Oils

Both natural and synthetic cutting oils, that is, oils which are mixedwith water and used as coolants and lubricants in, for example, latheand other metal-working operations, are extremely susceptible tomicrobial attack when formulated for use. This microbial attack leads tothe production of odor and to the eventual breakdown of the oilformulation. The isothiazolones of the invention are extremely useful incontrolling microbial build-up, thus preventing this deleteriousmicrobial attack, when incorporated in an emulsion or other diluted useformulation, either before or after the emulsification, at a level ofabout 10 to 10,000 parts per million of the use formulation. Thepreferred range of incorporation is about 20 to about 2000 parts permillion in the final emulsion.

In order to evaluate the isothiazolones as preservatives, samples ofcontaminated cutting oil from actual use operations were obtained. Thesecontaminated samples were used to inoculate uncontaminated cutting oilformulations to which an isothiazolone had been added. After one month,the formulation was examined to determine whether the isothiazolone waseffective in controlling microbial attack, and, if so, the formulationwas recontaminated. This examination and recontamination procedure wasfollowed for a period of six months.

Three formulations were employed in testing the isothiazolones. In thefirst formulation, the isothiazolone was added to a 1:60 oil-wateremulsion of an emulsifiable cutting oil containing petroleum oil andspecial emulsifiers (commercially available as Gulf's Gulfout SolubleOil). The inoculum for the first formulation was a naturallycontaminated oil emulsion. The results of this test are summarized inTable VI, which lists the isothiazolones tested and the minimumconcentration (in parts per million) of the isothiazolone which iseffective in controlling microbial attack for six months.

                  TABLE VI                                                        ______________________________________                                        Emulsifiable Cutting Oil Preservatives                                                            Min. Effective Conc.                                         Isothiazolone    (ppm)                                                     ______________________________________                                        2-n-butyl-3-isothiazolone                                                                         100-250                                                   2-n-hexyl-3-isothiazolone                                                                         20-30                                                     2-n-octyl-3-isothiazolone                                                                         20-30                                                     5-chloro-2-methyl-3-isothiazolone                                                                 < 100                                                     2-benzyl-3-isothiazolone                                                                          20-30                                                     2-phenyl-3-isothiazolone                                                                           50-100                                                   2-cyclohexyl-3-isothiazolone                                                                      500                                                       2-(4-chlorobenzyl)-3-isothiazolone                                                                 50                                                       2-(3,4-dichlorobenzyl)-3-isothiazolone                                                            500                                                       2-methyl-4,5-dichloro-3-isothiazolone                                                              40                                                       ______________________________________                                    

In the second formulation, the isothiazolone was added to a 1:40oil-water dilution of a synthetic cutting oil made up oftriethanolamine, sulfonated red oils, sodium nitrate, an anionic freeacid of a complex organic phosphate ester, and water (commerciallyavailable as H. Miller's Hamikleer No. 1591 Soluble Oil). The inoculumfor the second formulation was a naturally contaminated soluble oilemulsion to which was added Pseudomonas oleoverans one of the mosttroublesome contaminants of cutting fluid emulsions. The results of thistest are summarized in Table VII, which lists the isothiazolones testedand the minimum concentration (in parts per million) of theisothiazolone which is effective in controlling microbial attack for twomonths. In the third formulation, the isothiazolone was added at 0.5% tothe Hamikleer No. 1591 Soluble Oil prior to dilution with water. Theinoculum was a naturally contaminated soluble oil to which Pseudomonasoleoverans was added. The oil was then diluted to a 1:40 oil-water useformulation, in which the concentration of the isothiazolone was 125ppm. After one month, the formulation was examined to determine thenumber of bacteria surviving per milliliter of the emulsion. The resultsof this test are summarized in Table VII.

                  TABLE VII                                                       ______________________________________                                        Synthetic Cutting Oil Preservatives                                                            Min. Effect.                                                                              Bacteria/ml                                         Isothiazolone Conc; (ppm) (after 1 mo)                                     ______________________________________                                        2-n-butyl-3-isothiazolone                                                                      ≦ 25 0                                                2-n-hexyl-3-isothiazolone                                                                      100-250     0                                                2-benzyl-3-isothiazolone                                                                       50          0                                                None                         7,600,000                                        ______________________________________                                    

By way of comparison, it was determined that 500 ppm ofethylhexyldimethylalkylammonium cyclohexylsulfamate, a commercialcutting oil preservative, was ineffective in controlling bacterialattack in the Hamikleer cutting oil formulation.

The data from the above tests indicates the great' effectiveness of theisothiazolones of the invention as preservatives, when incorporated incutting oil formulations.

EXAMPLE B Control of Algae, Bacteria, Fungi, and Slime in Cooling Towers

A constantly recurring problem in water-cooling systems is theout-of-control growth of algae, bacteria, and fungi, and the associatedformation of slime. The isothiazolones of the invention are quiteeffective in controlling algae, bacteria, fungi, and slime formation inwater-cooling systems, and especially in water-cooling towers. When usedas a cooling tower microbicide, the isothiazolone will generally beadded at a concentration of about 1/2 to 1000 parts per million. Thepreferred concentration range of the isothiazolone is about 1 to 250parts per million.

The following test was employed to evaluate the isothiazolones ascooling tower microbicides.

A series of laboratory cooling towers, operated concurrently, was usedfor the evaluation study. The cooling tower liquid was an inorganicsalts medium, designed to promote algal development. Approximately 8liters of this liquid were held in the reservoir, at 35° C., andcontinuously recirculated over pine wood slats at a rate of 2.3 litersper minute. A similar, but more dilute medium was pumped continuouslyinto each cooling tower reservoir at a rate of 150 ml./hr. This wassufficient to replenish the volume lost in evaporation and to providethe system with a bleed-off of approximately 8 liters in 3 days. Aconstant volume was maintained in the reservoir by use of an over-flowdrain.

The cooling towers were heavily inoculated at the beginning of the studyand twice per week thereafter with microorganisms (algae, bacteria, andfungi) obtained from a large commercial cooling tower. If necessary, thefirst two weeks of operation were used to establish an adequatepopulation of microorganisms on the surfaces and in the reservoir, and,in addition, to determine volume losses due to evaporation. Whenstarting with a fouled tower, treatment was initiated by the addition ofthe isothiazolone at double the intended dose. When starting with aclean tower, treatment was initiated by the addition of theisothiazolone at the maintenance dose. The isothiazolone was then addedperiodically, usually once a week, to the diluting systems to restorethe concentration levels to the intended maintenance dose. Four factorswere considered in evaluating the effectiveness of the isothiazolenes ascooling-water microbicides-control of algae and control of non-algalslime as reflected in the appearance of the tower, and general controlof bacteria and fungi in the water itself. Three isothiazolones weretested by the above procedure and found to be effective at the indicatedminimum concentration in parts per million. These isothiazolones were2-n-hexyl-3-isothiazolone, at 5 ppm, 2-n-octyl-3-isothiazolone, at 5ppm, and 5-chloro-2-methyl-3-isothiazolone, at 2.5 ppm. Thehydrochloride salt of 5 chloro-2-methyl-3-isothiazolone was also foundto be effective.

In a supplementary test, isothiazolones were evaluated in controllingbacterial and fungal populations in water samples taken from actualcommercial cooling towers. The water sample from the cooling tower wasemployed as the diluent in preparing the dosage series of theisothiazolone being tested, and each diluted sample was examined forviable microorganisms 48 hours after the dilution. The minimum dose atwhich a zero plate count was obtained was considered to be the minimuminhibitory concentration for each isothiazolone tested. Under actual useconditions, a certain bacterial count might be tolerated, however, ifthe microbicide tested can keep the count from increasing and thus avoidslime formation. The results of this test are summarized in Table VIII,which lists the isothiazolones tested, and the average minimuminhibitory concentration (in parts per million) for each isothiazolone.

                  TABLE VIII                                                      ______________________________________                                        Water Treatment Microbicides                                                                      Min. Inhibitory Conc.                                        Isothiazolone    (ppm)                                                     ______________________________________                                        2-n-octyl-3-isothiazolone                                                                         500                                                       2-n-decyl-3-isothiazolone                                                                          37                                                       2-benzyl-3-isothiazolone                                                                          188                                                       4,5-dichloro-2-methyl-3-isothiazolone                                                             188                                                       ______________________________________                                    

The above tests demonstrate the effectiveness of the isothiazolones ofthe invention as water-cooling system microbicides.

EXAMPLE C Paint Film Mildewcides

Paint films from water-based and oil-based paints are quite susceptibleto mildewing, especially when applied on exterior surfaces. Presently,various mercury compounds are generally used as paint mildewcides.However, those mercury compounds have several disadvantages, includingtheir susceptibility to sulfide staining and their toxicity. Theisothiazolones of the invention are quite effective as paint mildewcideswithout the disadvantages of the mercurial mildewcides, and often withbetter performance than the mercury compounds. The concentration ofisothiazolone which is added to the paint can vary over a wide rangedepending on such factors as the type of paint involved, the locality ofapplication, and the type of surface on which the paint is applied.Generally, about 1/10 lb to 20 lb. of isothiazolone per 100 gallons ofpaint will be effective. The preferred range of incorporation is about1/2 lb. to 12 lb. of isothiazolone per 100 gallons of paint.

In order to evaluate isothiazolones as paint mildewcides, wood stickswere painted with water-based paint formulations to which theisothiazolone being tested had been added. The paint films were allowedto dry for two days, treated with a test fungus (Aspergillus niger orAspergillus cryzae) and after 7 to 20 days were examined for evidence ofmildew formation. The results of these tests are summarized in Table IX,which lists the isothiazolones tested and the minimum concentration (inpounds of isothiazolone per 100 gallons of paint) of isothiazolone whichinhibits the growth of the fungus.

                  TABLE IX                                                        ______________________________________                                        Paint Film Mildewcides                                                                            Min. Mildew Protection                                       Isothiazolone    Level (lb./100 gal.)                                      ______________________________________                                        2-n-butyl-3-isothiazolone                                                                         0.5-2                                                     2-n-hexyl-3-isothiazolone                                                                         0.5                                                       2-n-octyl-3-isothiazolone                                                                         0.25-2                                                    2-n-decyl-3-isothiazolone                                                                         4                                                         2-benzyl-3-isothiazolone                                                                          10                                                        5-chloro-2-methyl-3-isothiazolone                                                                 < 0.25                                                    2-cyclohexyl-3-isothiazolone                                                                      0.5-1                                                     4,5-dichloro-2-methyl-3-isothiazolone                                                             0.25                                                      5-chloro-2-benzyl-3-isothiazolone                                                                 > 0.25                                                    ______________________________________                                    

In addition to the above tests, 2-n-butyl-3-isothiazolone,2-n-hexyl-3-isothiazolone, 2-n-octyl-3-isothiazolone, and2-n-decyl-3-isothiazolone are found to be equally effective when testedin paint films from oil-based paints. Panels painted with paintscontaining 2-n-butyl-3-isothiazolone, 2-n-hexyl-3-isothiazolone, and2-n-octyl-3-isothiazolone were field tested and showed good to excellentresistance to mildew attack after at least six months of outdoorexposure.

The above tests and data demonstrate the excellent qualities of theisothiazolones when used as paint film mildewcides.

EXAMPLE D Preservatives for Vinyl or Acrylic Emulsion PolymerDispersions

On storage, aqueous dispersions of vinyl or acylic emulsion polymers,such as those used in making water-based paints, may be subject to abuildup of microorganisms which may lead to the production of odor ordiscoloration in the dispersion or to actual physical or chemicalbreakdown of the polymer.

Examples of such polymer dispersions include polyvinyl acetate;polyisobutylene; polystyrene; polymers of dienes, such as of isoprene,chloroprene, butadiene, including copolymers of butadiene with styrene,acrylonitrile or mixtures thereof; copolymers of (a) a soft acrylate,such as a (C₁ -C₈) alkyl acrylate (especially) methyl acrylate, ethylacrylate, butyl acrylate, 2-ethylhexyl acrylate or mixtures thereof),with (b) at least one hard comonomer, such as methyl methacrylate,acrylonitrile, styrene, vinyltoluene, vinyl acetate, and vinyl chloride,and (c) about 0.5 to 8% by weight of an α,β-monoethylenicallyunsaturated acid, such as acrylic, methacrylic, crotonic, or itaconicacid such as those described in Conn et al U.S. Pat. No. 2,795,564, June11, 1957; and blends of any of these polymer dispersions with each otheror with similar polymers containing a polar group, such as any of theblends mentioned in Scott U.S. Pat. No. 3,356,627, Dec. 5, 1967.

The isothiazolones of the invention are extremely effective incontrolling build-up of microorganisms in such aqueous polymerdispersions and in water-base paints made from them thus preventingdeleterious contamination of the dispersion or paint. An advantage ofusing the isothiazolones as preservatives is that in properconcentration they will also function as mildewcides after the polymerdispersion or paint has been used to make a coating or film. Whenemployed as preservatives, the isothiazolones are usually incorporatedin the polymer dispersion in a concentration range of about 1/2 to10,000 parts per million. The preferred concentration range is about 1to 2000 parts per million.

In order to evaluate the isothiazolones as preservatives for polymerdispersions, various samples of different types of emulsion polymerdispersions, to which an isothiazolone had been added, were contaminatedwith an inoculum consisting of naturally contaminated dispersion of thesame type or with a culture of organisms isolated from naturallycontaminated dispersions of the same type. For a period of six months,the samples were examined monthly for microbial contamination and thenreinoculated with fresh inoculum.

The isothiazolones were evaluated as preservatives in several differenttypes of commercially available acrylic polymer dispersions, of the typedescribed in the Conn et al and Scott U.S. patents mentioned above.Among the isothiazolones which were tested in one or more of suchacrylic polymer dispersions and found to give control of microbialactivity at a concentration of less than 1000 ppm in the dispersion are2-n-butyl-3-isothiazolone, 2-n-hexyl-3-isothiazlone,2-benzyl-3-isothiazolone, and 5-chloro-2-methyl-3-isothiazolone. In someof the dispersions, these isothiazolones resulted in control ofmicrobial activity at a concentration of less than 100 ppm.

EXAMPLE E Laundry Sanitizers

In order to sanitize the wash water, to kill bacteria in dirty clothes,and to prevent transfer of bacteria from dirty to clean clothes, it isdesirable to incorporate a sensitizing ingredient into laundry soaps anddetergents. The isothiazolones of the invention have been found to beuseful as laundry sanitizing agents. A wide concentration range can beemployed in formulating detergents or soaps with the isothiazolones ofthe invention as sanitizing agents. Generally, about 0.01 to 10% of theisothiazolone will be added to the soap or detergent, and the preferredrange is about 0.05 to 5%. The isothiazolone can also be added directlyto the wash water, generally at a concentration of about 1/2 to 1000parts per million.

In order to evaluate the isothiazolones of the invention as laundrysanitizing agents, several tests were employed. Initially, a timesurvival test was undertaken, employing the following procedure:

One milliliter of inoculum (the growth from one 24 hour slant suspendedin 20 ml. of phosphate buffer, using Escherichia coli and Staphylococcusaureus as the test organisms) is added to 99 ml. of test solution (a0.25% solution of a synthetic laundry detergent formulation containingthe desired concentration of the compound under test) and the flaskagitated to insure thorough mixing. Exactly 5 minutes, 10 minutes and 15minutes following inoculation, one ml. aliquots of the inoculated testsolution are removed from the test flask and placed in sterile 9 ml.neutralizing blanks. The blanks are agitated vigorously to assurecomplete homogenity and the resulting solution is then plated at 0, 10²,10³ and 10⁴ dilutions with Tryptone-Glucose-Extract Agar containingadditional neutralizer. All of the preceding steps were run at 25° C.The plates are incubated at 37° C. for 48 hours and then observed fortotal number of colonies. The results of the colony counts are recordedas surviving bacteria per ml. of original test solution, taking intoconsideration the dilution factor of the neutralizing blank and thedilution represented by the countable plate. The results of the timesurvival to test are summarized in Table X.

                  TABLE X                                                         ______________________________________                                        Time Survival Test                                                                        MIn. Effective Conc. (ppm)                                           Isothiazolone                                                                            5 Fin. exp.                                                                             10 min exp.                                                                             15 min. exp.                                ______________________________________                                        2-n-octyl-3-isothiazolone                                                                   75        50        10-25                                       2-n-decyl-3-isothiazolone                                                                   75        25        10-25                                       ______________________________________                                    

The isothiazolones were then evaluated for fabric sanitization activityand for wash water sanitization effectiveness.

A series of tests under actual cold water home laundry conditions wasconducted in an automatic washer using 12 gallons of cold water, a coldwater rinse, and a medium load of 4 lbs. of turkish towels as anequivalent to a typical home laundry load. The detergent product used inthe washer tests was a typical linear alkylate sulfonate (LAS) detergentformulation.

The operation schedule of the machine employed is as follows:

    ______________________________________                                                 Accumulated Time                                                                              Time in Minutes                                      Operation                                                                              in Minutes      For Each Operation                                   ______________________________________                                        fills    0-4             4                                                    washes    4-18           14                                                   stops    18-19           1                                                    empties  19-20           1                                                    spins    20-22           2                                                    fills    22-26           4                                                    rinses   26-28           2                                                    stops    28-29           1                                                    empties  29-30           1                                                    spins    30-32           2                                                    ______________________________________                                    

The detergent formulation was employed at a concentration in the washwater of 0.25%. The isothiazolone being tested was added to the washwater at a level of 50 ppm. The turkish bath towels used in the testswere pre-washed, dried and sterilized before use. Swatches of cottonTest Fabric 400 (2,3" swatches; 6, 11/2" swatches; 4 or 8, 1-7/8"diameter circular swatches) had been stapled to each of the towels ineach of the total of 10 runs made in the washing machine.

In a typical run, 113.5 gms. of the detergent (0.25%) were added to 12gallons of cold wash water (15°-17° C.) using a four pound turkish towelballast. The isothiazolone was added by removing 450 ml. of wash waterand replacing it with 450 ml. of anhydrous alcohol containing 2.271grams compound. This gave a concentration of 50 ppm of the isothiazolonein the wash water. The swatch towel to be inoculated was not yet added.The machine was agitated for one minute. As a check for bacterialcontent before inoculation, one milliliter of solution was removedaseptically from the washer and placed into a sterile 9 ml. neutralizerblank containing 0.5% sodium thioglycollate solution. Theswatch-containing towel was then inoculated with 4 ml. of a 24 hr.culture of S. aureus in AATCC Broth-3 ml. of inoculum were pipetted ontoone of the 3" swatches and 1 ml. onto one of the 11/2" swatches. Afterplacing the inoculated towel in the washer, the machine was restartedand one milliliter aliquots of wash water were removed aseptically atvarious points during the wash and rinse cycles. Each 1 ml. aliquot wasplaced into a sterile 9 ml. neutralizer blank containing 0.5% sodiumthioglycollate. These samples were then plated withTryptone-Glucose-Extract Agar containing 0.025% sodium thioglycollateand incubated for 48 hrs. at 37° C. A count was also made on the S.aureus inoculum to determine how many organisms would be introduced perml. of wash water.

The cotton swatches of Test Fabric-400 were removed aseptically from thetowel. The 3" inoculum swatch as well as the other 3" swatch were eachplaced into 100 ml. of sterile 0.5% sodium thioglycollate solution,shaken vigorously for one minute and the elution was plated in duplicatein Tryptone-Glucose-Extract Agar containing 0.025% sodiumthioglycollate.

The 11/2" inoculum swatch as well as another 11/2" swatch were placed insterile Petri dishes and covered with Tryptone Glucose Extract Agarcontaining 0.05% sodium thioglycollate. All the plates were incubatedfor 48 hours at 37° C.

The remaining 11/2" swatches as well as the circular swatches wereplaced in sterile Petri dishes and retained for testing by modified testmethods AATCC 90-1965 T and AATCC 100-1965 T.

The procedure used in other runs was the same as that above, whilevarying the inoculum and the isothiazolone tested.

The results of the wash water sanitization studies, the washed fabricswatch elutions, the plated washed fabric swatches and the residualactivity of the washed fabric swatches will be found in Tables XI andXII.

Table XI summarizes the results of the evaluation of2-n-decyl-3-isothiazolone and Table XII summarizes the results of theevaluation of 2-n-octyl-3-isothiazolone.

The isothiazolones were also tested to determine the antibacterialresidual which they imparted to fabrics.

Desized Test Fabric-400 was treated with aqueous solutions of thevarious isothiazolones under evaluation according to the tentative AOACLaboratory Treatment Method (Spindle Type). The actual treatment wascarried out in an Atlas Launderometer for 5 minutes at 25° C. using a1:10 fabric to treatment solution ratio.

Evaluation of the effectiveness of the antibacterial residuals impartedto the fabric was carried out by means of the tentative AOACModification of AATCC Methods 90-1965 T and 100-1965 T (Technical Manualof the American Association of Textile Chemists and Colorists, Volume44, 1968, pages B-175 to B-178). AATCC Method 90-1965 T measures the"bacteriostatic" residual and AATCC Method 100-1965 T measures the"self-sanitizing" residual in the fabric.

The results of the evaluations are given in Table XIII. It will be notedthat all three isothiazolones were able to impart a bacteriostaticresidual against both S. aureus and E. coli and a self-sanitizingresidual against S. aureus in the absence of a rinse followingtreatment. When two fresh water rinses were employed subsequent totreatment, the fabric treated with 2-n-decyl-3-isothiazolone alsoretained its "self-sanitizing" residual against S. aureus.

                                      TABLE XI                                    __________________________________________________________________________    Evaluation of 2-n-decyl-3-isothiazolone                                       __________________________________________________________________________    A.                                                                              Wash Water Sanitization Study                                               (1) Detergent Alone at 0.25% (No Bactericide)                                     (Number of bacteria/ml. of wash water)                                    Schedule for Removal of      E. coli                                          Wash Water Samples                                                                          S. aureus                                                                          E. coli                                                                            % kill.sup.1                                                                        S. aureus                                                                          % kill.sup.1                               __________________________________________________________________________    wash water before inoc.                                                                     0/ml 0/ml      0/ml                                             1 minute after inoc.                                                                         70  110,000   82,000                                           3 minute after inoc.                                                                        120  119,000   88,000                                           5 minute after inoc.                                                                        100  131,000   97,000                                           7 minute after inoc.                                                                        140  108,000   116,000                                          10 minute after inoc.                                                                       100  100,000   92,000                                           End of Wash cycle (14 min.)                                                                 160  99,000                                                                             0    96,000                                                                              0                                          __________________________________________________________________________    Start of Rinse cycle                                                                         0     450      860                                             End of Rinse cycle                                                                           40   5800      6100                                            __________________________________________________________________________    (2) Detergent at 0.25% Plus 2-n-decyl-3-isothiazolone at 50 ppm                   (Number of bacteria/ml. of wash water)                                    Schedule for Removal of      E. coli                                          Wash Water Samples                                                                           S. aureus                                                                         E. coli                                                                            % kill.sup.1                                                                        S. aureus                                                                          % kill.sup.1                               __________________________________________________________________________    wash water before inoc.                                                                      0      0      0                                                1 minute after inoc.                                                                        170  96,000    70,000                                           3 minute after inoc.                                                                         60  90,000    87,000                                           5 minute after inoc.                                                                        160  80,000    78,000                                           7 minute after inoc.                                                                        120  52,000    65,000                                           10 minute after inoc.                                                                       100  29,000    28,000                                           End of Wash cycle (14 min.)                                                                  90  17,200                                                                             82.1 12,900                                                                              85.1                                       __________________________________________________________________________    Start of Rinse cycle                                                                         0      70        30                                            End of Rinse cycle                                                                           20    520                                                                              91.0   500 91.8                                       __________________________________________________________________________     .sup.1 % kill represents the kill over and above that exhibited by            detergent alone  -                                                       

    B.                                                                              Washed Fabric Swatches (Elution)                                                               Number of Surviving Bacteria/3 inch Swatch                 Wash Solution  Inoculum                                                                          Inoculum Swatch                                                                          Uninoculated Swatch                             __________________________________________________________________________    Detergent alone                                                                             S. aureus                                                                           2,000     0                                               Detergent alone                                                                             E. coli                                                                            52,000     5600                                            Detergent and isothiazolone                                                                 S. aureus                                                                           1,500     0                                               Detergent and isothiazolone                                                                 E. coli                                                                            19,500     0                                               C.                                                                              Washed Fabric Swatches (Plated)                                                                Approximate Number of Organisms/Plate                      Wash Solution Inoculum                                                                           Inoculum Swatch                                                                          Uninoculated Swatch                             __________________________________________________________________________    Detergent alone                                                                             E. coli                                                                            >10,000    110                                             Detergent and isothiazolone                                                                 E. coli                                                                               262      4                                              __________________________________________________________________________

                                      TABLE XII                                   __________________________________________________________________________    Evaluation of 2-n-octyl-3-isothiazolone                                       __________________________________________________________________________    (A)                                                                              Wash Water Sanitization Study                                              (1)   Detergent Alone at 0.25% (No Bactericide)                                    (No. of bacteria/ml. of wash water)                                      Schedule for Removal of                                                       Wash Water Samples                                                                             S. aureus                                                                             E. coli                                                                              % kill.sup.1                                  __________________________________________________________________________    wash water before inoc.                                                                         0      0/ml.                                                1 minute after inoc.                                                                           220     90,000                                               3 minute after inoc.                                                                           450     92,000                                               5 minute after inoc.                                                                           480     96,000                                               7 minute after inoc.                                                                           530     73,000                                               10 minute after inoc.                                                                          690     86,000                                               End of Wash Cycle (14 min.)                                                                    1140    91,000 0                                             __________________________________________________________________________    Start of Rinse Cycle                                                                            0        120                                                End of Rinse Cycle                                                                             190       4630                                               __________________________________________________________________________    (2)  Detergent at 0.25% plus 2-n-octyl-3-isothiazolone                             (No. of bacteria 1 ml. of wash water)                                    Schedule for Removal of                                                       Wash Water Samples                                                                              S. aureus                                                                            E. coli                                                                              % kill.sup.1                                  __________________________________________________________________________    wash water before inoc.                                                                         40        0                                                 1 minute after inoc.                                                                           240     92,000                                               3 minute after inoc.                                                                           380     85,000                                               5 minute after inoc.                                                                           540     96,000                                               7 minute after inoc.                                                                           510     69,000                                               10 minute after inoc.                                                                          850     46,000                                               End of Wash Cycle (14 min.)                                                                    700     13,000 85.9                                          __________________________________________________________________________    Start of Rinse Cycle                                                                            2         10                                                End of Rinse Cycle                                                                             200       290  93.7                                          __________________________________________________________________________     (B)                                                                             Washed Fabric Swatches (Elution)                                                             Number of Surviving Bacteria/3 inch swatch                  Wash Solution                                                                              Inoculum                                                                           Inoculum Swatch                                                                          Uninoculated Swatch                              __________________________________________________________________________    Detergent Alone                                                                             E. coli                                                                           106,000    2300                                             Detergent & isothiazolone                                                                  E. coli                                                                             16,000      0                                              __________________________________________________________________________    (C)                                                                              Washed Fabric Swatches (Plated)                                                              Approximate Number of Organisms/Plate                       Wash Solution                                                                              Inoculum                                                                           Inoculum Swatch                                                                          Uninoculated Swatch                              __________________________________________________________________________    Detergent Alone                                                                            E. coli                                                                            >10,000    81                                               Detergent & isothiazolone                                                                  E. coli                                                                               400      3                                               __________________________________________________________________________     .sup.1 % kill represents the kill over and above that exhibited by            detergent alone                                                          

                                      TABLE XIII                                  __________________________________________________________________________    Antibacterial Residuals                                                                                     Modified AATCC Method                                                                    100-1965 T                                           Theor. Conc.  90-1965 T  No. of Surviving S. aureus                  Conc. of Active                                                                        of ACt. Bacteri-                                                                      Rinses                                                                              Diameter of Zone of                                                                      Cells per Fabric Sample                     Bactericide in                                                                         cide on Following                                                                           Inhibition in mm.                                                                        Zero time                                                                           After 22 hrs.                  Isothiazolone                                                                        Treatment Soln.                                                                        Fabric  Treatment                                                                           S. aureus                                                                           E. coli                                                                            count Exposure                       __________________________________________________________________________    None   --       --      None  no zone                                                                             no zone                                                                            150,00                                                                              >100,000,000                   2-n-hexyl-3-                                                                  isothiazolone                                                                        50 ppm.  500 ppm.                                                                              None  1.5-2.5                                                                             1.0-1.5    C                              2-n-cetyl-3-                                                                  isothiazolone                                                                        50 ppm.  500 ppm.                                                                              None  1.0-4.0                                                                             trace      5,500                          2-n-decyl-3-                                                                  isothiazolone                                                                        50 ppm.  500 ppm.                                                                              None  10    1.0-4.0    0                              __________________________________________________________________________

Several other isothiazolones were tested by the AATCC Methods 90-1965 Tand 100-1965 T, referred to above, and found to give a bacteriostatic ora self-sanitizing residual on fabric against S. aureus and/or E. coli1000 ppm or less. Among these isothiazolones were2-n-butyl-3-isothiazolone, 2-n-dodecyl-3-isothiazolone, and5-chloro-2-methyl-3-isothiazolone.

The above tests and data demonstrate the usefulness of theisothiazolones as laundry sanitizing agents.

EXAMPLE F Use as Bacteriostats and Mildewstats for Paper and PaperProducts

Biocides are incorporated into paper and paper products for the purposeof imparting an antimicrobial residual to control growth of fungi(mildew) and bacteria. The isothiazolones of the invention have beenfound to be useful as bacteriostats and mildewstats for paper and paperproducts. When used as paper bacteriostas and mildewstats, theisothiazolones are generally applied to the paper in aqueous solution bya conventional technique such as dipping or spraying. The3-isothiazolone can have a wide concentration in the solution and willgenerally be about 0.1 to about 10,000 parts per million by weight, andpreferably about 10 to about 1000 parts per million. Depending on theamount of solution uptake by the paper, the concentration ofisothiazolone in the treated paper will usually range from about 0.2 toabout 30,000 parts per million, and preferably about 25 to 3,000 partsper million by weight.

Isothiazolones were screened for this activity as follows:

Pieces of Whatman No. 1 filter paper were dipped momentarily insolutions containing either 1,000 ppm, 100 ppm, or 10 ppmisothiazolones. The treated papers were air dried, cut into 11/2 inchsquares, and tested for resistance to bacterial and mildew organismsaccording to Military Medical Specifications No. 7-864-790.

The results, shown in Table XIV, show that isothiazolone treatmentsimpart bacterial and fungal protection to the paper.

                  TABLE XIV                                                       ______________________________________                                        Isothiazolones as Paper Bacteriostats and Mildewstats                                              Bacteriostatic                                                                            Mildewstatic                                                      Results     Results                                                           Zone of     Growth of                                              PPM active Inhibition vs.                                                                            Chaetomium                                   Compound  in Treatment                                                                             S. aureus, mm                                                                             globosum.sup.a                               Tested    Solution   Rep. 1  Rep. 2                                                                              Rep. 1                                                                              Rep. 2                               ______________________________________                                        2-n-butyl-4-                                                                            1000       0-5     0-5   0     0                                    isothiazolin-                                                                           100        0       0     0     0                                    3-one     10         0       0     3+    3+                                   2-n-hexyl-4-                                                                            1000       13      12    0     0                                    isothiazolin-                                                                           100        trace   trace 0     0                                    3-one     10         0       0     0     0                                    2-n-octyl-4-                                                                            1000       12      12    0     0                                    isothiazolin-                                                                           100        trace   0     0     0                                    3-one     10         0       0     0     0                                    2-t-octyl-4-                                                                            1000       1-2     1-2   0     0                                    isothiazolin-                                                                           100        0       0     0     0                                    3-one     10         0       0     3+    3+                                   2-benzyl-4-                                                                             1000       8-9     6-9   0     0                                    isothiazolin-                                                                           100        0       0     3+    2+                                   3-one     10         0       0     4+    4+                                   Control   0          0       0     4+    4+                                   (No. Cmpd.)                                                                   ______________________________________                                         .sup.a 0 = No Growth (complete inhibition)                                    1+  = trace of growth                                                         2+  =  slight growth                                                          3+ = moderate growth                                                          4+ = heavy growth (no inhibition)                                        

EXAMPLE G Mildewstats for Leather

Mildewcides are used to prevent mildew damage of stored leather,especially during processing. Isothiazolones of the invention have beenfound to be useful as mildewstats for leather. When used as leathermildewstats, the isothiazolones are generally applied to the wet-tannedstock in aqueous solution by a conventional technique such as dipping orspraying. The concentration of the isothiazolone in the treatmentsolution can be varied over a wide concentration range, and willgenerally be about 0.1 to about 10,000, and preferably about 1 to about1000, parts per million by weight. The solution can be applied to theleather at any convenient rate, but will commonly be applied at 100%float, that is, at a rate of 100 grams of treatment solution per 100grams of wet-tanned stock.

Isothiazolone-treated chrome-tanned leather samples were evaluated forresidual fungistatic activity by means of the American ChemistAssociation's Mold Resistance test (L-1). The results, presented inTable XV, show that isothiazolones are effective in this application.

                  TABLE XV                                                        ______________________________________                                        Mildew Resistance of Chrome-Tanned Leather                                    Treated with Isothiazolones                                                                      ALCA-L-1 Test Results                                                         Mildew Growth After                                        Compound  PPM      Treatment 1    2    3    4                                 Tested    Cmpd,.sup.a                                                                            Rep. No.  wk.  wks. wks. wks.                              ______________________________________                                        2-n-octyl-4-                                                                            10       1         0    0     1+   2+                               isothiazolin-      2         0    0     1+   2+                               3-one                                                                                   25       1         0    0    0    0                                                    2         0    0    0    0                                           50       1         0    0    0    0                                                    2         0    0    0    0                                           100      1         0    0    0    0                                                    2         0    0    0    0                                 2-n-hexyl-4-                                                                            10       1         0    0     1+   2+                                                  2         0    0     2+   3+                                         25       1         0    0    0    0                                                    2         0    0    0    0                                           50       1         0    0    0    0                                                    2         0    0    0    0                                           100      1         0    0    0    0                                                    2         0    0    0    0                                 ______________________________________                                         .sup.a Treatment concentration based on wet weight of chrome stock; 100%      float.                                                                   

EXAMPLE H Application of Mildewcides for Wood Surfaces, Hard Surfaces,and Fabric

Mildewcides are used to control growth of mildew, and subsequentdevelopment of mildew odor, discoloration and/or deterioration of woodsurfaces, ceramic (hard) surfaces, and fabric in humid environments.Isothiazolones of the invention have been found to be useful in thesemildewcidal applications. When used for the surface treatment of wood,ceramic, brick, and fabric, the isothiazolones are generally applied tothe material to be treated in aqueous or organic solvent solution byconventional methods in a sufficient amount to saturate the surface, andthe treated material can then be dried. The concentration of theisothiazolone in the treatment solution can be varied over a wideconcentration range, and will generally be about 0.1 to about 10,000,and preferably about 1 to about 1000, parts per million by weight.

Isothiazolones were found to be effective in tests recommended by theU.S. Environmental Protection Agency for efficacy evaluations ofproducts intended for mildew control on fabric, hard surfaces (ceramictiles) and wood surfaces.

The tests were performed as follows:

(1) Fabric Test:

The test fungi used in this procedure were Aspergillus niger (A. nigerATTC 6275) and Penicillium glaucum, both cultures were maintained onEmmon's agar. Spores were harvested after 8-10 days incubation andsuspended in 0.85% sodium chloride and 0.2% Triton X-100 surfactant.Test spore suspensions containing 5 million spores/ml of each funguswere mixed 50:50 and sprayed on the test fabric strips (8-20 ouncecotton duck, 1"×3") after chemical treatment.

Chemical treatment of test strips (10 replicates/treatment) wasaccomplished by immersing fabric for three minutes in test solution.After drying overnight at 37° C., each strip was sprayed with sporeinoculum and hung in an 8 ox. screw-capped jar containing 1 inch ofsterile water. Observations for fungus growth were made for 4 weeks.

(2) Hard Surface Test (Ceramic Tiles)

An A. niger spore suspension (5 million/ml.) diluted 20:1 withCzapek-Dox nutrient solution was used to inoculate ceramic tiles treatedas indicated for fabrics. The dried inoculated tiles were placed on thesurface of sterile agar in a petri dish and incubated at 25° C. for 7days.

(3) Wood Block Test

The test fungus was A. niger and the inoculum prepared as in the abovesection. Ponderosa pine sapwood blocks (2.5×1.5 cm with hole drilled inone corner) were immersed in treatment solution for 3 minutes, air driedfor several hours, and then spray inoculated, hung in jars over water aswith the fabric, and incubated at 22°-30° C. for 4 weeks.

The results of these tests, which are summarized in Tables XVI, XVII,and XVIII, show that the isothiazolones are effective as mildewcides inthese applications.

                  TABLE XVI                                                       ______________________________________                                        Mildew Protection of Fabric Strips                                                       Effective Level.sup.1 (PPM)                                        Test Compound                                                                              1 wks.   2 wks.   3 wks. 4 wks.                                  ______________________________________                                        2-n-octyl-4-                                                                  isothiazolin-                                                                 3-one        5        5        5      5                                       5-chloro-2-                                                                   methyl-4-isothia-                                                             zolin-3-one  5        5        5      5                                       Sodium penta-                                                                 chlorophenate                                                                              250      250      250    250                                     ______________________________________                                         .sup.1 Min. effective level to protect 10 replicates.                         .sup.2 Incubation period after treatment.                                

                  TABLE XVII                                                      ______________________________________                                        Mildew Protection of Hard Surfaces                                            (Ceramic Tiles)                                                                                 Minimum effective level                                                       to protect 10 replicates                                    Test Compound     after 7 days Incubation, PPM                                ______________________________________                                        2-n-octyl-4-                                                                  isothiazolin- -3-one                                                                            5-10                                                        5-chloro-2-methyl-                                                            4-isothiazolin-                                                               3-one             >100                                                        Sodium pentachlorophenate                                                                       10,000                                                      ______________________________________                                    

                  TABLE XVIII                                                     ______________________________________                                        Mildew Protection of Wood Surfaces                                                        Minimum Effective Level                                                       (PPM) to protect                                                  Test        10 replicate Wood Blocks                                          Compound    1 wk.      2 wks.  3 wks.  4 wks.                                 ______________________________________                                        2-n-octyl-4-                                                                              100 (or less)                                                                            100      200    200                                    isothiazolin-                                                                 3-one                                                                         5-chloro-2-                                                                   methyl-4-                                                                     isothiazolin-                                                                              10        100     100-1000                                                                             100-1000                                3-one                                                                         (HCl salt).sup.1                                                              Sodium penta-                                                                 chlorophenate                                                                             100        1000    1000   1000                                    ______________________________________                                         .sup.1 2 Replicates in test.                                             

EXAMPLE I Wood Preservative Application

The isothiazolones of the invention have also been found to be effectivein controlling microbial attack on wood. For use as wood preservatives,the isothiazolones can be applied to the wood in aqueous or organicsolvent solution by any convenient conventional technique. Generally,the isothiazolone will be applied to give a retention of about 0.001 toabout 10, and preferably about 0.005 to about 5, pounds of isothiazoloneper cubic foot of wood.

To determine the value of isothiazolones as wood preservatives, the ASTMLaboratory Soil-Block Culture Test (D-1413-61) was performed. Theresults in Table XIX show wood preservative activity of2-n-octyl-3-isothiazolone in terms of decreased percentage weight lossof isothiazolone treated wood compared to control without test compoundin toluene treatment solution

                  TABLE XIX                                                       ______________________________________                                        Isothiazolones as Wood Preservatives                                          PPM       Retention                                                           2-n-octyl-4-                                                                            pounds     Percentage Wt. loss                                      isothiazolin-                                                                           isothiazolone                                                                            of Block                                                 3-one     per cubic  vs. Lentinus                                                                              vs. Lenzites                                 in toluene                                                                              foot of wood                                                                             lepideus culture                                                                          trabea culture                               ______________________________________                                        0         0          44.6        62.7                                         8         0.002      35.3        38.1                                         17        0.005      25.5        56.4                                         33        0.009      26.1        54.9                                         67        0.019      21.4        32.8                                         134       0.038      8.4         17.2                                         267       0.076      2.8         1.7                                          ______________________________________                                    

EXAMPLE J Fungicidal Disinfectant

In the process of chemical disinfection, a fungicidal agent can be usedto kill the fungus contaminants associated with disease. To demonstrateefficacy in this application, isothiazolones were evaluated in theA.O.A.C. Fungicidal Test.

The results presented in Table XX, which shows the maximum effectivedilution vs. Trichophyton interdigitale-ATCC 9533, the causative agentfor athelete's foot, demonstrate further fungicidal effectiveness ofisothiazolones of the invention.

                  TABLE XX                                                        ______________________________________                                        Evaluation of Isothiazolones Against                                          Trichophyton interdigitals                                                                        Maximum Effective                                         Compound            Dilution (PPM)                                            ______________________________________                                        2-n-butyl-4-isothiazolin-3-one                                                                    1:800 (1250 PPM)                                          2-n-hexyl-4-isothiazolin-3-one                                                                    1:1500 (600 PPM)                                          2-n-octyl-4-isothiazolin-3-one                                                                    1:1000 (1000 PPM)                                         2-t-octyl-4-isothiazolin-3-one                                                                    1:800 (1250 PPM)                                          2-n-nonyl-4-isothiazolin-3-one                                                                    >1:1200 (<830 PPM)                                        2-n-dodecyl-4-isothiazolin-3-one                                                                  >1:1200 (<830 PPM)                                        2-cyclohexyl-4-isothiazolin-3-one                                                                 >1:1200 (<830 PPM)                                        2-methyl-4,5-dichloro-4-                                                      isothiazolin-3-one  1:800 (1250 PPM)                                          ______________________________________                                    

EXAMPLE K Cosmetic Preservatives

Microbicides are used as preservatives to prevent the growth ofmicroorganisms in cosmetic formulations and to kill microbialcontaminants introduced during manufacturing and during use of theproduct. Isothiazolones of the invention have been found to beefficacious in this application. When used in cosmetic formulations, theconcentration of isothiazolone can be varied greatly depending on theparticular formulation, and will generally be about 1 to about 20,000,and preferably about 10 to about 3,000, parts per million by weight.

To evaluate the effectiveness of isothiazolones of the invention ascosmetics preservatives, the following standard cosmetic formulationswere prepared:

    ______________________________________                                        Formulation No. 1                                                             cetyl alcohol, 20%                                                            mineral oil, 20%                                                              sorbitan monooleate, 0.5%                                                     polyoxyethylene sorbitol monooleate, 4.5%                                     deionized water, 55%                                                          Formulation No. 2                                                             mineral oil 24.5%                                                             sorbitan monostearate, 1.5%                                                   polyoxyethylene soritan monostearate, 8.5%                                    deionized water, 65.5%                                                        ______________________________________                                    

Samples (50 g) containing varying concentrations of isothiazolones wereinoculated with a pooled suspension of the following microorganisms:

Pseudomonas oleoverans

Escherichia coli

Staphylococcus aureus

Serrotia sp.

Aspergillus sp.

Penicillium sp.

Streptomyces sp.

Saccharomyces cerevisiae

Cladosporium resinae

and incubated at room temperature. The samples were re-inoculatedmonthly for 8 months to simulate contamination during use of the product(Table XXII) or were inoculated only initially, to simulatecontamination during manufacturing (Table XXI). Effectiveness was basedon the absence of viable microorganisms, as determined by variousstandard microbiological techniques.

The results of these tests, shown in Tables XXI and XXII, show theeffectiveness of isothiazolones of the invention in preventing thegrowth of microorganisms in cosmetic preservatives.

                                      TABLE XXI                                   __________________________________________________________________________    Isothiazolones as Cosmetic Preservatives                                      in Formulations Contaminated During Preparation                                                     Minimum Effective Concentration (ppm)                                         Formulation No.                                         Compound              1         2                                             __________________________________________________________________________    2-cyclohexyl-4-isothiazolin-3-one                                                                   1000 (2).sup.1                                                                          1000 (2)                                      2-n-butyl-4-isothiazolin-3-one                                                                      100 (8)   1000 (8)                                      2-n-cyclopropyl-4-isothiazolin-3-one                                                                500 (8)   500 (8)                                       2-methyl-4-isothiazolin-3-one (HCl)                                                                 250 (8)   500 (8)                                       2-n-butyl-5-chloro-4-isothiazolin-3-one                                                             100 (8)   50 (8)                                        2-n-propyl-5-chloro-4-isothiazolin-3-one                                                            50 (8)    50 (8).sup.2                                  2-ethyl-5-chloro-4-isothiazolin-3-one                                                               50 (8)    50 (8).sup.2                                  2-methyl-5-chloro-4-isothiazolin-3-one (HCl)                                                        50 (8)    50 (8)                                        2-methyl-4-bromo-5-chloro-4-isothiazolin-3-one                                                      50 (8)    500 (8)                                       2-phenyl-5-chloro-4-isothiazolin-3-one                                                              250 (8)   1000 (6)                                      formaldehyde          >1000 (1) 1000 (6)                                      methyl p-hydroxybenzoate                                                                            >3000 (1) >3000 (1)                                     propyl p-hydroxybenzoate                                                                            >3000 (1) >3000 (1)                                     __________________________________________________________________________     .sup.1 (Months) from inoculation                                              .sup.2 failure vs. fungi at 1000 ppm                                     

                                      TABLE XXII                                  __________________________________________________________________________    Isothiazolones of Cosmetic Preservatives                                      in Formulations Re-contaminated Monthly                                                             Minimum Effective Concentration (ppm)                                         Formulation No.                                         Compound              1         2                                             __________________________________________________________________________    2-n-butyl-4-isothiazolin-3-one                                                                      250 (8).sup.1                                                                           250 (8)                                       2-n-cyclopropyl-4-isothiazolin-3-one                                                                500 (8)   500 (8)                                       2-methyl-4-isothiazolin-3-one (HCl)                                                                 500 (8)   500 (8)                                       2-n-butyl-5-chloro-4-isothiazolin-3-one                                                             100 (8)   50 (8)                                        2-n-propyl-5-chloro-4-isothiazolin-3-one                                                            50 (8)    50 (8)                                        2-ethyl-5-chloro-4-isothiazolin-3-one                                                               50 (8)    50 (8)                                        2-methyl-5-chloro-4-isothiazolin-3-one (HCl)                                                        50 (8)    50 (8)                                        2-methyl-4-bromo-5-chloro-4-isothiazolin-3-one                                                      50 (8)    >1000 (8)                                     2-phenyl-5-chloro-4-isothiazolin-3-one                                                              500 (8)   >1000 (<8)                                    Methyl p-hydroxybenzoate                                                                            >3000 (<8)                                                                              >3000 (<8)                                    __________________________________________________________________________     .sup.1 (Months) from initial inoculation                                 

EXAMPLE L Soap Bacteriostat Application

Bacteriostats and bactericides are employed in soap, as a means tocontrol bacterial microflora on the skin. The isothiazolones of theinvention have been found to be effective as bacteristatic andbactericidal additives to soap. While the concentration of theisothiazolone in the soap can be varied greatly depending on the soapformulation, the isothiazolone used, the degree of activity desired, andrelated factors, it will generally be present at about 0.01 to about 3%,and preferably about 0.1 to about 3% by weight.

Isothiazolones were evaluated for this application using a serialdilution test and the published Protein Adsorption Test of Bechtold andLawrence (Proceeding of the Scientific Section of the Toilet GoodsAssociation, 24, December, 1955).

The test used in these evaluations was a serial dilution test similar tothat described above for Table III, except dilutions were made also inthe presence of soap. Biological compatibility of isothiazolone withsoap was indicated in this test when the maximum bacteriostatic dilutionof the isothiazolone was not adversely affected by the presence of soap.The results are shown in Table XXIII.

The Protein Adsorption Test of Bechtold and Lawrence shows the potentialof soap bacteriostat candidates in terms of adsorption. As shown inTable XXIV, adsorption was indicated by the presence of a zone ofinhibition.

                  TABLE XXIII                                                     ______________________________________                                        Serial-Dilution                                                               Test for Soap Bacteriostats                                                                     Maximum                                                                       Bacteriostatic.sup.1                                                          Dilution                                                    Compound            Without Soap                                                                             With Soap                                      ______________________________________                                        2-n-octyl-4-isothiazolin-3-one                                                                    1:128,000  1:128,000                                      2-n-decyl-4-isothiazolin-3-one                                                                    1:1,000,000                                                                              1:1,000,000                                    2-n-dodecyl-4-isothiazolin-3-one                                                                  1:1,000,000                                                                              1:1,000,000                                    2-methyl-5-chloro-4-isothiazolin-3-one                                                            1:128,000  1:128,000                                      2,2'-dihydroxy-3,5,6-3',5',6'-                                                hexachlorodiphenyl methane                                                    (Hexachlorophene)   1:2,000,000                                                                              1:2,000,000                                    ______________________________________                                         .sup.1 vs. Staph. Aureus (ATCC 6538)                                          .sup.2 Ivory Flakes, 50:1 ratio soap to compound                         

                  TABLE XXIV                                                      ______________________________________                                        Protein Adsorption                                                            Soap Bacteriostat Test                                                                                 Average                                                                       Zone of                                              Compound                 Inhibition, mm.sup.1                                 ______________________________________                                        2-n-octyl-4-isothiazolin-3-one                                                                         trace                                                2-n-dodecyl-isothiazolin-3-one                                                                         trace                                                2-isopropyl-5-chloro-4-isothiazolin-3-one                                                              15.5                                                 2-n-butyl-4-bromo-4-isothiazolin-3-one                                                                 1.0                                                  2-n-hexyl-4-bromo-5-chloro-4-isothiazolin-3-one                                                        8.5                                                  2-n-hexyl-4-bromo-4-isothiazolin-3-one                                                                 2.0                                                  2-n-octyl-5-chloro-4-isothiazolin-3-one                                                                7.0                                                  2-(4-chlorophenyl-)-5-chloro-4-isothiazolin-3-one                                                      1.3                                                  2-cyclohexyl-4,5-dichloro-4-isothiazolin-3-one                                                         7.0                                                  Hexachlorophene          8.0                                                  No compound              0                                                    ______________________________________                                         .sup.1 zone diameter minus diameter of protein film disc                 

The above data demonstrate the effectiveness of isothiazolones of theinvention as soap bacteriostats.

The isothiazolones were also found to be quite useful as microbicides infiber spin finish formulation, such as nylon spin finish formulations.In evaluating the isothiazolones in this application, samples ofnaturally contaminated spin finish formulations were obtained and usedto inoculate uncontaminated formulations to which an isothiazolone hadbeen added. After a one month incubation, the test formulations wereexamined for contamination, and then recontaminated. In one such test,2-benzyl-3-isothiazolone was found to be very effective in controllingmicrobial contamination in fiber spin finish formulations at aconcentration of 250 parts per million or less.

It is to be understood that changes and variations may be made withoutdeparting from the spirit and scope of the invention as defined by theappended claims.

We claim:
 1. A cosmetic formulation which comprises a cosmetic agent, asurfactant, and a fungicidally or a bactericidally effective amount of acompound of the formula ##STR14## wherein Y is an unsubstituted orsubstituted alkyl, alkenyl, or alkynyl group of 1 to 18 carbon atoms, anunsubstituted or substituted cycloalkyl group having a 3 to 6 carbonatom ring and up to 12 carbon atoms, an unsubstituted or substitutedaralkyl group of up to 10 carbon atoms, or an unsubstituted orsubstituted aryl group of up to 10 carbon atoms,R is hydrogen, halogen,or a (C₁ -C₄)alkyl group,and R' is hydrogen, halogen, or a (C₁ -C₄)alkylgroup, or a salt of a compound of the above formula with a strong acid.2. The formulation of claim 1 wherein Y is (1) an unsubstituted alkylgroup of 1 to 18 carbon atoms, (2) a substituted alkyl group having atleast one hydrogen atom replaced by hydroxy, halo, cyano, alkylamino,dialkylamino, phenylamino, halophenylamino, carboxy, carbalkoxy, alkoxy,aryloxy, or carbamoxy, wherein the total number of carbon atoms in thesubstituted alkyl group does not exceed 18, (3) an unsubstituted orhalo-substituted alkenyl group of 2 to 18 carbon atoms, (4)unsubstituted or halo-substituted alkynyl group of 2 to 28 carbon atoms,(5) an unsubstituted or alkyl-substituted cycloalkyl group having a 3 to6 carbon atom ring and up to 12 carbon atoms, (6) an unsubstituted or ahalo-, lower alkyl-, or lower alkoxy-substituted aralkyl group whereinthe total number of carbon atoms in the aralkyl group does not exceed10, or (7) an unsubstituted or halo-, nitro-, lower alkyl-, or lowercarbalkoxy-, substituted aryl group wherein the total number of carbonatoms in the aryl group does not exceed 10, R is hydrogen, halogen, or a(C₁ -C₄) alkyl group, and R' is hydrogen, halogen, or a (C₁ -C₄)alkylgroup or a salt of a compound of the above formula with a strong acid.3. The formulation of claim 2 in which R is hydrogen or halogen, and R'is hydrogen or halogen, or a salt of a compound of the above formulawith a strong acid.
 4. The formulation of claim 3 in which Y is anunsubstituted alkyl or cycloalkyl group, R' is chlorine, and R ishydrogen.
 5. The formulation of claim 4 in which Y is an alkyl group.